Aesthetic design of skin-sparing mastectomy incisions for immediate autologous tissue breast reconstruction in asian women.
The advent of skin-sparing mastectomy has allowed for the reconstruction of the breast and nipple with improved cosmesis. However, the nipple-areolar complex (NAC) in Asian patients is more pigmented and scars easily. Therefore, commonly described incisions tend to result in poor aesthetic outcomes in Asian patients with breast cancer. We describe an algorithmic approach to skin-sparing mastectomy incisions in Asian patients on the basis of the location of the biopsy scar and the tumor site and size. Four incision types are described: peri-areolar, a peri-areolar incision with a second distant skin paddle, "racquet handle," and peri-areolar with adjacent skin excision. 281 immediate breast reconstructions were performed between May 2001 and February 2012 after skin-sparing mastectomy. The mastectomy incisions used included the peri-areolar design (n=124, 44%), peri-areolar design with a second distant skin paddle (n=39, 14%), "racquet handle" (n=21, 7.5%), and peri-areolar design with adjacent skin excision (n=42, 14%). The traditional elliptical incision and other variants where the NAC outline was not preserved were performed in the remaining 55 patients. The average follow-up was 44.7 months during which there was 1 case of total flap loss and 7 cases of partial flap necrosis; all remaining flaps survived. 24% of the patients (68/281) underwent subsequent nipple reconstruction. Our algorithm avoids breast incisions that are randomly placed or excessively long and prevents the unnecessary sacrifice of normal breast skin. This allows skin-sparing mastectomy and immediate breast reconstruction to be performed with a consistently achievable aesthetic result in Asian women without neglecting oncological safety.


[Liposome phospholipid substitution and lung function in surfactant deprived rats].
In vivo activity of an artificial surfactant was studied in surfactant depleted rats. After tenfold alveolar lavage, PaO2, tidal volume, and compliance of the respiratory system fell to one third of initial value. Substitution of large unilamellar vesicles containing 90% Dipalmitoylphosphatidylcholine and 10% unsaturated phosphatidylglycerol largely restored oxygenation and lung mechanics in most animals. Complete normalization with weaning from the ventilator, however, was achieved neither with liposomes nor with natural surfactant concentrate.


Preservation of kidney function with combined inhibition of NADPH oxidase and angiotensin-converting enzyme in diabetic nephropathy.
Antihypertensive therapies such as angiotensin-converting enzyme-1 inhibitors (ACEi) slow the decline in renal function seen with diabetic nephropathy, although there is still progression ultimately to end-stage renal disease. The aim of this study was to determine if there were added renoprotective benefits seen by combining ACEi with blockade of NADPH oxidase. Sprague-Dawley diabetic and non-diabetic rats were randomized to receive intervention therapy with apocynin (15 mg/kg/day, weeks 16-32), apocynin + the ACEi ramipril (1 mg/kg/day, weeks 16-32), or ramipril alone (1 mg/kg). All three treatments retarded the development of albuminuria in the diabetic rats. Apocynin conferred its benefit either as a monotherapy or in combination with ramipril without affecting blood pressure per se. Renal morphological injury was attenuated by all three treatment strategies. Diabetes was associated with increasing renal fibronectin and type IV collagen protein expression, with the combination regimen resulting in the highest decrease in extracellular matrix accumulation. All three treatments prevented the diabetes-associated increases in renal cytosolic superoxide generation as well as urinary isoprostanes. While renal TGF-beta1 activation was reduced by ramipril treatment but not by apocynin as a monotherapy, kidney cortical membranous VEGF was reduced by apocynin as monotherapy and dual therapy but not by ramipril alone. Combination of NADPH oxidase blockade with ACE inhibitors is a promising regimen which warrants further investigation as a way to confer additional renoprotection in diabetes.


Production of anti-Helicobacter pylori urease-specific immunoglobulin in egg yolk using an antigenic epitope of H. pylori urease.
The potential therapeutic effects of Helicobacter pylori-specific immunoglobulin (IgY-Hp) derived from egg yolk and identification of the immunodominant H. pylori proteins have previously been reported. In this study, the urease epitope that is recognized by IgY-Hp was identified and used as an immunogen to produce urease-specific IgY (IgY-HpU). Epitope regions were mapped and peptides of selected epitope regions were synthesized. The IgY-Hp titre against synthetic peptides was evaluated using ELISA analysis. Hens were immunized with synthetic peptides conjugated with BSA. Urease activity was quantified by measuring the optical density of an indicator dye. Of the five synthetic peptides assayed, a peptide representing 15 amino acid residues of UreB (UB-3; aa 396-410, DNDNFRIKRYLSKYT) was specifically recognized by the IgY-Hp. Immunization of hens with BSA-conjugated UB-3 resulted in the generation of IgY-HpU. IgY-HpU markedly reduced H. pylori urease activity by 80 % as compared to control IgY (IgY-BSA). The availability of the synthetic UreB-derived peptide enabled the production of highly specific anti-urease IgY, which had a significant inhibitory effect on H. pylori urease activity. Therefore, specific IgY-HpU produced using the synthetic peptide may be an effective tool against infection by H. pylori.


Session Introduction: TOWARDS ETHICAL BIOMEDICAL INFORMATICS: LEARNING FROM OLELO NOEAU, HAWAIIAN PROVERBS.
Innovations in human-centered biomedical informatics are often developed with the eventual goal of real-world translation. While biomedical research questions are usually answered in terms of how a method performs in a particular context, we argue that it is equally important to consider and formally evaluate the ethical implications of informatics solutions. Several new research paradigms have arisen as a result of the consideration of ethical issues, including but not limited for privacy-preserving computation and fair machine learning. In the spirit of the Pacific Symposium on Biocomputing, we discuss broad and fundamental principles of ethical biomedical informatics in terms of Olelo Noeau, or Hawaiian proverbs and poetical sayings that capture Hawaiian values. While we emphasize issues related to privacy and fairness in particular, there are a multitude of facets to ethical biomedical informatics that can benefit from a critical analysis grounded in ethics.


Synaptic relationships of a type of GABA-immunoreactive neuron (clutch cell), spiny stellate cells and lateral geniculate nucleus afferents in layer IVC of the monkey striate cortex.
The precise stimulus specificity of striate cortical neurons is strongly influenced by processes involving gamma-aminobutyric acid (GABA). In the visual cortex of the monkey most afferents from the lateral geniculate nucleus terminate in layer IVC. We identified a type of smooth dendritic neuron (clutch cell) that was immunoreactive for GABA, and whose Golgi-impregnated dendrites and axon were largely restricted to layer IVC beta. The slightly ovoid somata were 8-12 micron by 12-15 micron and the dendritic field was often elongated, extending 80-200 micron in one dimension. The axonal field was 100-150 micron in diameter and densely packed with large bulbous boutons. Although mainly located in IVC beta both the dendritic and axonal processes entered IVC alpha. Fine structural features of GABA-immunoreactive and-impregnated clutch cells and impregnated spiny stellate cells were compared. Clutch cells had more cytoplasm, more densely packed mitochondria and endoplasmic reticulum, and made type II as opposed to type I synapses. A random sample of 159 elements postsynaptic to three clutch cells showed that they mainly terminated on dendritic shafts (43.8-58.5%) and spines (20.8-46.3%), rather than somata (10-17%). The majority of the postsynaptic targets were characteristic of spiny stellate cells. This was directly demonstrated by studying synaptic contacts between an identified GABA positive clutch cell and the dendrites and soma of an identified spiny stellate cell. The termination of clutch cells mainly on dendrites and spines of spiny stellate cells suggests that they interact with other inputs to the same cells. Following an electrolytic lesion in the ipsilateral lateral geniculate nucleus we examined the distribution of degenerating terminals on three identified spiny stellate neurons in layer IVC beta. Out of eight synapses from the lateral geniculate nucleus one was on a dendritic shaft, the rest on spines. Only a small fraction of all synapses on the cells were from degenerating boutons. A clutch cell within the area of dense terminal degeneration was not contacted by terminals from the lateral geniculate nucleus. The results show that layer IVC in the monkey has a specialized GABAergic neuron that terminates on spiny stellate cells monosynaptically innervated from the lateral geniculate nucleus. Possible functions of clutch cells may include inhibitory gating of geniculate input to cortex; maintenance of the antagonistic subregions in the receptive fields; and the creation from single opponent of double colour opponent receptive fields.


Risk of thrombocytopenic, haemorrhagic and thromboembolic disorders following COVID-19 vaccination and positive test: a self-controlled case series analysis in Wales.
There is a need for better understanding of the risk of thrombocytopenic, haemorrhagic, thromboembolic disorders following first, second and booster vaccination doses and testing positive for SARS-CoV-2. Self-controlled cases series analysis of 2.1 million linked patient records in Wales between 7th December 2020 and 31st December 2021. Outcomes were the first diagnosis of thrombocytopenic, haemorrhagic and thromboembolic events in primary or secondary care datasets, exposure was defined as 0-28 days post-vaccination or a positive reverse transcription polymerase chain reaction test for SARS-CoV-2. 36,136 individuals experienced either a thrombocytopenic, haemorrhagic or thromboembolic event during the study period. Relative to baseline, our observations show greater risk of outcomes in the periods post-first dose of BNT162b2 for haemorrhagic (IRR 1.47, 95%CI: 1.04-2.08) and idiopathic thrombocytopenic purpura (IRR 2.80, 95%CI: 1.21-6.49) events; post-second dose of ChAdOx1 for arterial thrombosis (IRR 1.14, 95%CI: 1.01-1.29); post-booster greater risk of venous thromboembolic (VTE) (IRR-Moderna 3.62, 95%CI: 0.99-13.17) (IRR-BNT162b2 1.39, 95%CI: 1.04-1.87) and arterial thrombosis (IRR-Moderna 3.14, 95%CI: 1.14-8.64) (IRR-BNT162b2 1.34, 95%CI: 1.15-1.58). Similarly, post SARS-CoV-2 infection the risk was increased for haemorrhagic (IRR 1.49, 95%CI: 1.15-1.92), VTE (IRR 5.63, 95%CI: 4.91, 6.4), arterial thrombosis (IRR 2.46, 95%CI: 2.22-2.71). We found that there was a measurable risk of thrombocytopenic, haemorrhagic, thromboembolic events after COVID-19 vaccination and infection.


Teaching diabetic ptients about self-management.
It takes approximately 20 min to teach a patient how to use Dextrostix reagent strips in conjunction with an Eyetone reflectance meter (or the more recently introduced Glucochek). Blood glucose monitoring is no more difficult or time consuming than conventional urine testing and can be learned and executed by patients of various levels of intelligence. It is important to emphasize the need for an adequate drop of blood and the use of Monolet blood lancets has greatly facilitated this. Emphasis is put on the most useful times to make recordings and the importance of good record-keeping. The more intelligent patients readily learn how to adjust their insulin to improve their blood glucose readings while the less intelligent need help and advice. Experience has shown that no only are blood glucose levels improved but also hypoglycemic episodes are less commmon.


Biomimetic Chaotic Sensor for Moderate Static Magnetic Field.
The effects of a static magnetic field on systems with chaotic dynamical behavior have attracted little attention so far. Here, Chua's electronic circuit with an inductor placed in a static uniform magnetic field operating in a chaotic double-scroll regime is studied experimentally. The effect of the magnetic field on the duty cycle factor and the spike count rate, with spikes defined by crossings between the scrolls of the double-scroll attractor, is described. A slow monotonic variation in the duty cycle factor and constant spike count rate is observed for magnetic field intensities up to the threshold, where both these metrics change severely; the dynamic trajectory remains on one scroll and spikes disappear. The dependence of the static magnetic field intensity on Chua's circuit resistivity at the threshold is given. Two biomimetic magnetic chaotic sensors are proposed: one based on one Chua's circuit and another that can have various transfer functions and is composed of several independent Chua's circuits.


Macrophages in resistance to candidiasis.
Candida albicans, an increasingly common opportunistic pathogenic fungus, frequently causes disease in immunodeficient but not immunocompetent hosts. Clarifying the role of the phagocytic cells that participate in resistance to candidiasis not only is basic to understanding how the host copes with this dimorphic pathogen but also will expedite the development of innovative prophylactic and therapeutic approaches for treating the multiple clinical presentations that candidiasis encompasses. In this review, we present evidence that a diverse population of mononuclear phagocytes, in different states of activation and differentiation and from a variety of host species, can phagocytize C. albicans blastoconidia via an array of opsonic and nonopsonic mechanisms and can kill C. albicans blastoconidia and hyphae by means of oxygen-dependent and -independent mechanisms. Reactive nitrogen intermediates should now be added to the well-established candidacidal reactive oxygen intermediates of macrophages. Furthermore, what were thought to be two independent pathways, i.e., nitric oxide and superoxide anion, have now been shown to combine to form a potent macrophage candidacidal molecule, peroxynitrite. In contrast to monocytes and neutrophils, which are important in resistance to early stages of C. albicans infections, more differentiated macrophages activated by cytokines such as gamma interferon participate in the acquired resistance of hosts with C. albicans-specific, cell-mediated immunity. Evidence presented in this review demonstrates that mononuclear phagocytes, in some instances in the absence of other professional phagocytes such as neutrophils, play an import role in resistance to systemic and mucosal candidiasis.


Thermal Response to High-Power Holmium Laser Lithotripsy.
The aim of this study was to investigate "caliceal" fluid temperature changes during holmium laser activation/lithotripsy using settings up to 40 W power output with different irrigation flow rates. The experimental system consisted of a glass test tube (diameter 10 mm/length 75 mm) filled with deionized water, to mimic a calix. Real-time temperature was recorded using a thermocouple (Physitemp, NJ) positioned 5 mm from the bottom of the tube. A 200 μm laser fiber (Flexiva; Boston Scientific, MA) was introduced through the working channel of a disposable ureteroscope (LithoVue; Boston Scientific) and the laser fiber tip was positioned 15 mm above the bottom of the test tube. Deionized water irrigation (room temperature) through the working channel of the ureteroscope was delivered at flow rates of 0, 7-8, 14-15, and 38-40 mL/minute. A 120-W holmium laser (pulse 120; Lumenis, CA) was used. The following settings were explored: 0.5 J × 10 Hz, 1.0 J × 10 Hz, 0.5 J × 20 Hz, 1.0 J × 20 Hz, 0.5 J × 40 Hz, 1.0 J × 40 Hz, and 0.5 J × 80 Hz. During each experiment, the laser was activated continuously for 60 seconds. Temperature increased with increasing laser power output and decreasing irrigation flow rate. The highest temperature, 70.3°C (standard deviation 2.7), occurred with laser setting of 1.0 J × 40 Hz and no irrigation after 60 seconds of continuous laser firing. None of the tested laser settings and irrigation parameters produced temperature exceeding 51°C when activated for only 10 seconds of continuous laser firing. High-power holmium settings fired in long bursts with low irrigation flow rates can generate high fluid temperatures in a laboratory "caliceal" model. Awareness of this risk allows urologist to implement a variety of techniques (higher irrigation flow rates, intermittent laser activation, and potentially cooled irrigation fluid) to control and mitigate thermal effects during holmium laser lithotripsy.


Splenectomy in hereditary spherocytosis: Review of 1,657 patients and application of the pediatric quality indicators.
The objective of the present study is to profile the outcome and safety of pediatric patients undergoing splenectomy with hereditary spherocytosis (HS) using a nationwide sample and the Agency for Healthcare Research and Quality (AHRQ) Pediatric Quality Indicators (PDIs). A retrospective cross-sectional descriptive analysis of a non-overlapping combination of the National Inpatient Sample (NIS), and Kids' Inpatient Database (KID) databases (1988-2004) were performed. These combined databases contain information from nearly 93 million discharges in the United States. Children with an age at admission of <18 years of age and HS (ICD-9 diagnosis code of 282.0) who underwent total splenectomy (ICD-9 procedure code of 41.5) were identified. Variables of gender, race, co-existing diagnoses, hospital type, and charges adjusted to 2006 dollars, length of stay, inpatient mortality, and complications were collected. PDIs were identified for each patient by linking the data obtained from the NIS and KID databases with the PDIs using the AHRQ Quality Indicators Wizard. Splenectomy for HS was associated with low morbidity and mortality. Accompanying cholecystectomy and/or appendectomy appeared to be safely performed at the same operation. Of the 13 PDIs identified by AHRQ as potentially avoidable adverse events, none were observed to occur in more than 1% of the patients. Based on the results of this study, splenectomy in patients with HS appears safe and to result in a minimal number of potentially preventable complications as identified by the AHRQ PDIs. We have successfully demonstrated use of the indicators to aid in the analysis of a specific surgical procedure within a subset of the pediatric population.


Kytococcus aerolatus sp. nov., isolated from indoor air in a room colonized with moulds.
A Gram-positive, coccoid bacterial isolate (02-St-019/1(T)), forming beige pigmented colonies was obtained from an indoor air sample. Based on 16S rRNA gene sequence similarity studies it was determined that this isolate 02-St-019/1(T) belonged to the genus Kytococcus, showing sequence similarties of 98.6% to Kytococcus schroeteri DSM 13884(T) and 98.3% to Kytococcus sedentarius DSM 20547(T), respectively. The diagnostic diaminoacid of the peptidoglycan was lysine, cell wall sugars were ribose and xylose. The major menaquinones detected were MK-7 and MK-8. The polar lipid profile consisted of the major phospholipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine and phosphatidylinositol mannoside. Fatty acid patterns were composed of major amounts of the iso- and anteiso-branched fatty acids anteiso C(17:0), iso C(15:0) and iso C(17:0) and unsaturated fatty acids (C(17:1) omega8c, iso C(17:1) omega9c, and C(17:1) omega8c) with smaller amounts of the straight-chain fatty acids C(15:0), C(16:0) and C(17:0). The results of DNA-DNA hybridizations and physiological and biochemical tests clearly allowed a genotypic and phenotypic differentiation of strain 02-St-019/1(T) from the two described Kytococcus species. On the basis of these results a novel species to be named Kytococcus aerolatus sp. nov., is proposed, with the type strain 02-St-019/1(T) (=DSM 22179(T)=CCM 7639(T)).


Multiphase mixed-effects models for repeated measures data.
Behavior that develops in phases may exhibit distinctively different rates of change in one time period than in others. In this article, a mixed-effects model for a response that displays identifiable regimes is reviewed. An interesting component of the model is the change point. In substantive terms, the change point is the time when development switches from one phase to another. In a mixed-effects model, the change point can be a random coefficient. This possibility allows individuals to make the transition from one phase to another at different ages or after different lengths of time in treatment. Two examples are reviewed in detail, both of which can be estimated with software that is widely available.


Deuterium-depleted water has stimulating effects on long-term memory in rats.
Deuterium-depleted water (DDW) is a water which has a 6-7-fold less concentration of the naturally occurring deuterium (20-25ppm vs. 150ppm). While administrated for a longer period, it may reduce the concentration of deuterium throughout the body, thus activating cellular mechanisms which are depending on protons (channels, pumps, enzyme proteins). The aim of the present work was to study, for the first time in our knowledge, the possible influence of deuterium-depleted water (DDW) chronic administration in normal Wistar rats, as compared to a control group which received distilled water, on spatial working memory and the locomotor activity (as studied through Y-maze) or both short-term and long-term spatial memory (assed in radial 8 arms-maze task). Our results presented here showed no significant modifications in terms of spatial working memory (assessed through spontaneous alternation percentage) and locomotor activity (expressed through the number of arm entries) in Y-maze, as a result of DDW ingestion. Also, no significant differences between the DDW and control group were found in terms of the number of working memory errors in the eight-arm radial maze, as a parameter of short-term memory. Still, we observed a significant decrease for the number of reference memory errors in the DDW rats. In this way, we could speculate that the administration of DDW may generate an improvement of the reference memory, as an index of long-term memory. Thus, we can reach the conclusion that the change between the deuterium/hydrogen balance may have important consequences for the mechanisms that govern long-term memory, as showed here especially in the behavioral parameters from the eight-arm radial maze task.


Chitosan-cellulose composite for wound dressing material. Part 2. Antimicrobial activity, blood absorption ability, and biocompatibility.
Chitosan (CS), a polysaccharide derived from chitin, the second most abundant polysaccharide, is widely used in the medical world because of its natural and nontoxic properties and its innate ability for antibacterial and hemostasis effects. In this study, the novel composites containing CS and cellulose (CEL) (i.e., [CEL + CS]), which we have previously synthesized using a green and totally recyclable method, were investigated for their antimicrobial activity, absorption of anticoagulated whole blood, anti-inflammatory activity through the reduction of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and the biocompatibility with human fibroblasts. The [CEL + CS] composites were found to inhibit the growth of both Gram positive and negative micro-organisms. For examples, the regenerated 100% lyophilized chitosan material was found to reduce growth of Escherichia coli (ATCC 8739 and vancomycin resistant Enterococcus faecalis (ATCC 51299) by 78, 36, and 64%, respectively. The composites are nontoxic to fibroblasts; that is, fibroblasts, which are critical to the formation of connective tissue matrix were found to grow and proliferate in the presence of the composites. They effectively absorb blood, and at the same rate and volume as commercially available wound dressings. The composites, in both air-dried and lyophilized forms, significantly inhibit the production of TNF-α and IL-6 by stimulated macrophages. These results clearly indicate that the biodegradable, biocompatible and nontoxic [CEL + CS] composites, particularly those dried by lyophilizing, can be effectively used as a material in wound dressings.


Mechanical properties and early clinical experience with xenograft biomaterials.
Evaluation of new materials for ligament and tendon reconstruction has opened up a new area of orthopaedic research. This report outlines the mechanical properties, animal evaluations, and early clinical results from tests of a xenograft biomaterial of bovine source treated with a gluteraldehyde-based process. A clinical trial under FDA guidelines is currently underway.


Discovery of a functional, contracted heme-binding motif within a multiheme cytochrome.
Anaerobic ammonium-oxidizing (anammox) bacteria convert nitrite and ammonium via nitric oxide (NO) and hydrazine into dinitrogen gas by using a diverse array of proteins, including numerous c-type cytochromes. Many new catalytic and spectroscopic properties of c-type cytochromes have been unraveled by studies on the biochemical pathways underlying the anammox process. The unique anammox intermediate hydrazine is produced by a multiheme cytochrome c protein, hydrazine synthase, through the comproportionation of ammonium and NO and the input of three electrons. It is unclear how these electrons are delivered to hydrazine synthase. Here, we report the discovery of a functional tetraheme c-type cytochrome from the anammox bacterium Kuenenia stuttgartiensis with a naturally-occurring contracted Cys-Lys-Cys-His (CKCH) heme-binding motif, which is encoded in the hydrazine synthase gene cluster. The purified tetraheme protein (named KsTH) exchanged electrons with hydrazine synthase. Complementary spectroscopic techniques revealed that this protein harbors four low-spin hexa-coordinated hemes with His/Lys (heme 1), His/Cys (heme 2), and two His/His ligations (hemes 3 and 4). A genomic database search revealed that c-type cytochromes with a contracted CXCH heme-binding motif are present throughout the bacterial and archaeal domains in the tree of life, suggesting that this heme recognition site may be employed by many different groups of microorganisms.


Adoption of an algorithm and client pathway for the aged and disabled: training implications for the health and human service sectors.
This paper reports on a state agency's training activities undertaken to totally redesign a long-term-care (LTC) delivery system as part of the national Aging and Disability Resource Center (ADRC) initiative. Through the development of an ADRC algorithm and the implementation of a corresponding client pathway, NJ DACS, a division of aging, aligned 14 separate core functions necessary for lifespan services. A Training Academy facilitated the adoption of five new health service delivery products and processes by state and county health and human services personnel. Intensive training activities resulted in the algorithm and client pathway framework being successfully disseminated in all 21 counties within a short timeframe. Barriers to training were reduced and acceptance of new protocols and processes were facilitated leading to rapid adoption. Implications for training of health and human service personnel are presented. Full adoption of the complete ADRC model across the state was directly linked to agency software integration. Promoting standardized service delivery for the aging population through the use of an algorithm and parallel client pathway is feasible as a training model for health care service delivery.


Transcriptome Analysis Reveals Candidate Genes for Cold Tolerance in Drosophila ananassae.
Coping with daily and seasonal temperature fluctuations is a key adaptive process for species to colonize temperate regions all over the globe. Over the past 18,000 years, the tropical species Drosophila ananassae expanded its home range from tropical regions in Southeast Asia to more temperate regions. Phenotypic assays of chill coma recovery time (CCRT) together with previously published population genetic data suggest that only a small number of genes underlie improved cold hardiness in the cold-adapted populations. We used high-throughput RNA sequencing to analyze differential gene expression before and after exposure to a cold shock in coldtolerant lines (those with fast chill coma recovery, CCR) and cold-sensitive lines (slow CCR) from a population originating from Bangkok, Thailand (the ancestral species range). We identified two candidate genes with a significant interaction between cold tolerance and cold shock treatment: GF14647 and GF15058. Further, our data suggest that selection for increased cold tolerance did not operate through the increased activity of heat shock proteins, but more likely through the stabilization of the actin cytoskeleton and a delayed onset of apoptosis.


Hyperphosphorylated tau Inflicts Intracellular Stress Responses That Are Mitigated by Apomorphine.
Abnormal phosphorylation of the microtubule-binding protein tau in the brain is a key pathological marker for Alzheimer's disease and additional neurodegenerative tauopathies. However, how hyperphosphorylated tau causes cellular dysfunction or death that underlie neurodegeneration remains an unsolved question critical for the understanding of disease mechanism and the design of efficacious drugs. Using a recombinant hyperphosphorylated tau protein (p-tau) synthesized by the PIMAX approach, we examined how cells responded to the cytotoxic tau and explored means to enhance cellular resistance to tau attack. Upon p-tau uptake, the intracellular calcium levels rose promptly. Gene expression analyses revealed that p-tau potently triggered endoplasmic reticulum (ER) stress, Unfolded Protein Response (UPR), ER stress-associated apoptosis, and pro-inflammation in cells. Proteomics studies showed that p-tau diminished heme oxygenase-1 (HO-1), an ER stress associated anti-inflammation and anti-oxidative stress regulator, while stimulated the accumulation of MIOS and other proteins. P-tau-induced ER stress-associated apoptosis and pro-inflammation are ameliorated by apomorphine, a brain-permeable prescription drug widely used to treat Parkinson's disease symptoms, and by overexpression of HO-1. Our results reveal probable cellular functions targeted by hyperphosphorylated tau. Some of these dysfunctions and stress responses have been linked to neurodegeneration in Alzheimer's disease. The observations that the ill effects of p-tau can be mitigated by a small compound and by overexpressing HO-1 that is otherwise diminished in the treated cells inform new directions of Alzheimer's disease drug discovery.


Antigen-specific T-helper cells stimulate H-2-compatible and H-2-incompatible B-cell blasts polyclonally.
Lipopolysaccharide (LPS)-activated B-cell blasts from C57BL/6J nu/nu spleen cells develop into IgM-secreting clones after stimulation by antigen-specific T-helper cells of C57BL/6J origin. Although induction of help is antigen-dependent, help itself acts polyclonally. 1 of 1--3 B-cell blasts is restimulated in a homologous fashion by LPS, or in a heterologous fashion by sheep erythrocyte (SRC)- or horse erythrocyte (HRC)-activated T-helper cells. The repertoire of activated B-cell blasts reflects the polyclonal nature of activation: approximately 1 in 1,000--3,000 restimulated B-cell blasts is specific for SRC, 1 in 300--1,000 is specific for HRC, and 1 in 100--300 specific for trinitrophenylated SRC (TNP30-SRC). B-cell blasts that are either H-2 compatible or H-2 incompatible with the antigen-activated T-cell help are stimulated polyclonally in similar high frequencies. Thus, neither antigen nor H-2 compatibility are required to stimulate a B-cell blast into the next cell cycle.


Histologic classification of non-small-cell lung cancer over time: reducing the rates of not-otherwise-specified.
The importance of histologic classification in selecting the appropriate systemic therapy for non-small-cell lung cancer (nsclc) came to attention in 2007. In British Columbia, that information was communicated through international and national meetings, our centralized cancer care program, and to the medical community at large in multidisciplinary forums. We examined the effects of those education programs on the categorization of nsclc and associated systemic treatment practices. The BC Cancer Agency provides cancer care to 4.6 million residents of British Columbia. A retrospective review of all stage iiib and ivnsclc patients referred in 2007 and 2011 collected baseline characteristics, treatment, and outcomes. Histology was classified using the International Classification of Diseases for Oncology, 3rd edition, for the Canadian Cancer Registry. In 2007, 671 patients were referred, and 170 received chemotherapy; in 2011, the relevant figures were 680 and 197 respectively. Baseline characteristics in the cohorts were not statistically significantly different in 2007 and 2011. Histologic classifications in 2007 were 41% nonsquamous, 13% squamous, and 46% not otherwise specified (nos); in 2011, they were 63%, 17%, and 20% respectively. Exposure to pemetrexed in any line of therapy in 2007 was 22% for nonsquamous, 17% for squamous, and 10% for nos; in 2011, exposure was 39%, 3%, and 37% respectively. Exposure to epidermal growth factor receptor tyrosine kinase inhibitor (egfrtki) in 2007 was 36%, 22%, and 33%; in 2011, it was 64%, 60%, and 63%. Median overall survival duration, 2007 versus 2011, was 3.25 months versus 3.57 months with best supportive care, and 11.31 months versus 11.54 months with chemotherapy. The specificity of nsclc histologic categorization improved in 2011 compared with 2007, with a reduction of 26 percentage points in the rate of nos disease. The proportion of patients treated with chemotherapy over time remained the same, but the use of pemetrexed and egfrtki increased. The increased accuracy in histologic classification resulted in more appropriate utilization of systemic drugs.


Nutritional consequences of radiotherapy in nasopharynx cancer patients.
Nasopharynx cancer (NPC) occurs frequently in southern China. Radiotherapy (RT) is the standard treatment for this cancer, and weight loss is commonly observed during and for a prolonged period after RT. The determinants of this phenomenon are not well known. The nutritional status of 38 NPC patients was assessed serially before and for a 6-mo period after RT. Body weight, body composition (by dual X-ray absorptiometry), basal metabolic rate (BMR, by indirect calorimetry), and calorie intake (by 3-day dietary record) were documented at pre-RT (T0), end-RT (T1), 2 mo post-end-RT (T2), and 6 mo post-end-RT (T3). The BMI at end-RT was 21.5 +- 3.7 kg/m2 (range = 13.7-27.9 kg/m2) and was significantly lower than that at pre-RT (P < 0.001). Body weight at T1-T3 was significantly lower than that at T0 (P < 0.001). Mean percentage weight loss was 10.8% at end-RT. Fifty-five percent of patients (20 of 30) had 10% weight loss by the end of RT. BMR corrected for body weight did not change significantly among the four time points, but BMRs corrected for lean body mass at T2 and T3 were significantly lower than that at T0 (P < 0.01). Patients' calorie intake generally declined from T0 to T1 (from 1,857 P < 411 kcal to 11,68 +- 549 kcal, P < 0.001) and then increased from T1 to T3. The calorie intakes at T2 and T3 were not significantly different from that at T0 but were significantly higher than that at T1. Patients were in negative energy balance before, during, and up until 6 mo after RT. The recovery in body weight lagged behind the recovery of dietary intake. Eighty-two percent of NPC patients had significant weight loss and was in negative energy balance at the end of RT, which persisted for at least 6 mo. This finding suggests that there is room for improvement in the nutritional status of patients with NPC treated with RT.


[Diagnostics and correction of vascular disorders in upper extremity muscular transplants].
The features of clinical presentation, diagnosis and treatment of the acute circulatory disorders of muscular transplants in 7 patients after 59 transplantations of revascularized and reinnervated muscle grafts in severe consequences of upper extremities injuries were presented in the article. Tissue complex on pedicle were transplanted to upper extremity in Volkmann contracture (n=47), in consequences of the gunshot wound (n=7) and electric trauma (n=5). Signs of circulatory disorders of the transplanted tissue complex were registered shortly after the operation in 7 patients. Diagnosis was based on the findings of visual and ultrasound investigations. In all cases of circulatory disorders disorders patients were reoperated within 24 hours. Arterial thrombosis occurred in 3 cases and venous - in 2. The cause of impaired venous outflow was a hematoma in 2 cases. Repeated reconstruction of anastomoses with wound drainage in 5 cases led to restoration of blood circulation in transplants. Reoperation was not successful in 2 cases with developing transplant necrosis.


Hemolysin of Mycoplasma pneumoniae: tentative identification as a peroxide.
Mycoplasma pneumoniae produces a soluble hemolysin active against guinea pig erythrocytes. This hemolysin appears to be a peroxide, since catalase or peroxidase inhibits its activity. The action of catalase and peroxidase is specific, since heating the enzymes abolishes their effect on the hemolysin. In addition, 3-amino- 1,2,4-triazole, a potent inhibitor of catalase, reverses the inhibitory effect of the enzyme. The hemolysin of M. laidlawii is also a peroxide. The hemolysins of M. pneumoniae and M. laidlawii seem unique for microbial organisms since the bacterial hemolysins which have been described have been protein or lipid in nature.


False-negative scintigraphy with Tc-99m sestamibi in stage IV neuroblastoma.
Tc-99m sestamibi, originally developed for myocardial studies, has been used as a tumor-seeking agent. Recently, the agent also was reported to be a functional tracer to predict multidrug resistance-related p-glycoprotein expression in tumor tissue. The current report presents the authors' experience with sestamibi tumor scintigraphy in a neuroblastoma. Although I-131 MIBG tumor imaging and Tc-99m MDP bone scanning accurately demonstrated the extent of the disease, Tc-99m sestamibi showed no accumulation in primary and metastatic foci. Lack of sestamibi uptake was initially thought to be suggestive of failure to respond to chemotherapy because of p-glycoprotein expression. However, the patient responded well to chemotherapy and complete remission was achieved. The failure of Tc-99m sestamibi to detect a neuroblastoma and the lack of sestamibi accumulation in the tumor may not always be related to chemotherapy resistance.


2D Wavelet-Scalogram Deep-Learning for Seizures Pattern Identification in the Post-Hypoxic-Ischemic EEG of Preterm Fetal Sheep.
Neonatal seizures after an hypoxic-ischemic (HI) event in preterm newborns can contribute to neural injury and cause impaired brain development. Preterm neonatal seizures are often not detected or their occurrence underestimated. Therefore, there is a need to improve knowledge about preterm seizures that can help establish diagnostic tools for accurate identification of seizures and for determining morphological differences. We have previously shown the superior utility of deep-learning algorithms for the accurate identification and quantification of post-HI microscale epileptiform transients (e.g., gamma spikes and sharp waves) in preterm fetal sheep models; before the irreversible secondary phase of cerebral energy failure starts by the bursts of high-amplitude stereotypic evolving seizures (HAS) in the signal. We have previously developed successful deep-learning algorithms that accurately identify and quantify the micro-scale transients, during the latent phase. Building up on our deep-learning strategies, this work introduces a real-time deep-learning-based pattern fusion approach to identify HAS in the 256Hz sampled post-HI data from our preterm fetuses. Here, for the first time, we propose a 17-layer deep convolutional neural network (CNN) classifier fed with 2D wavelet-scalogram (WS) images of the EEG patterns for accurate seizure identification. The WS-CNN classifier was cross-validated over 1812 manually annotated EEG segments during ~6 to 48 hours post-HI recordings. The classifier accurately recognized HAS patterns with 97.19% overall accuracy (AUC = 0.96).Clinical relevance-The promising results from this preliminary work indicate the ability of the proposed WS-CNN pattern classifier to identify HI-related seizures in the neonatal preterm brain using 256Hz EEG; the frequency commonly used clinically for data collection.


Neurosurgical materials and devices. Report on regulatory agencies and advisory groups.
The current status of voluntary consensus standards writing procedures in neurosurgery and the current progress of government efforts to regulate materials and devices are described. A survey of the national and international standards writing bodies is presented, along with an introduction to related organizations and agencies and nomenclature. The intent of this review is to provide the neurosurgeon with a reference source regarding past and present neurosurgical activities in the materials and devices field. When President Ford signed the 1976 Medical Device Amendments on May 28, 1976, the Food and Drug Administration assumed direct legal authority to control medical devices and potentially assumed the power to regulate those professionals using them.


Exercise Intervention Improves the Metabolic Profile and Body Composition of Southwestern American Indian Adolescents.
The Southwestern American Indian population carries a high prevalence of metabolic syndrome and obesity, placing this group at higher risk than the general population for developing early type 2 diabetes and cardiovascular disease, likely impacting overall lifespan. This study aims to evaluate the impact of early lifestyle interventions which promote healthy eating and regular exercise on risk factors contributing to the development of the metabolic syndrome among the adolescent Zuni Pueblo population. We describe a prospective, single site, community-based cohort study performed among sixty-five adolescent Zuni Indians aged 13.9 ± 1.7 years who were recruited between March 2011 and January 2014. The study intervention consisted of a targeted, tri-weekly exercise regimen with nutritional counselling, and the primary study outcomes included changes from baseline in metabolic profile (fasting lipids, A1c), vital signs (blood pressure, resting heart rate) and anthropometric characteristics of the study group. 41 participants have anthropometric data measured at baseline and after completion, biochemical data are available from 30 participants, and body composition data from 26 patients. Using the paired Student's t-test with Bonferroni correction, significant improvements were shown in pediatric BMI percentile, fasting lipid profile, A1C, total body fat, and fat free mass after six months of exercise and nutritional intervention. A simple, standardized fitness program among Southwest American Indian adolescents was effective at reducing fasting lipids and adiposity, as well as improving glycemic indices over the course of six months.


Temporal Dynamics of Bacterial Communities in Soil and Leachate Water After Swine Manure Application.
Application of swine manure to agricultural land allows recycling of plant nutrients, but excess nitrate, phosphorus and fecal bacteria impact surface and drainage water quality. While agronomic and water quality impacts are well studied, little is known about the impact of swine manure slurry on soil microbial communities. We applied swine manure to intact soil columns collected from plots maintained under chisel plow or no-till with corn and soybean rotation. Targeted 16S-rRNA gene sequencing was used to characterize and to identify shifts in bacterial communities in soil over 108 days after swine manure application. In addition, six simulated rainfalls were applied during this time. Drainage water from the columns and surface soil were sampled, and DNA was extracted and sequenced. Unique DNA sequences (OTU) associated with 12 orders of bacteria were responsible for the majority of OTUs stimulated by manure application. Proteobacteria were most prevalent, followed by Bacteroidetes, Firmicutes, Actinobacteria, and Spirochaetes. While the majority of the 12 orders decreased after day 59, relative abundances of genes associated with Rhizobiales and Actinomycetales in soil increased. Bacterial orders which were stimulated by manure application in soil had varied responses in drainage waters over the course of the experiment. We also identified a "manure-specific core" of five genera who comprised 13% of the manure community and were not significantly abundant in non-manured control soils. Of these five genera, Clostridium sensu stricto was the only genus which did not return to pre-manure relative abundance in soil by day 108. Our results show that enrichment responses after manure amendment could result from displacement of native soil bacteria by manure-borne bacteria during the application process or growth of native bacteria using manure-derived available nutrients.


Mechanical Ventilation and ARDS in the ED: A Multicenter, Observational, Prospective, Cross-sectional Study.
There are few data regarding mechanical ventilation and ARDS in the ED. This could be a vital arena for prevention and treatment. This study was a multicenter, observational, prospective, cohort study aimed at analyzing ventilation practices in the ED. The primary outcome was the incidence of ARDS after admission. Multivariable logistic regression was used to determine the predictors of ARDS. We analyzed 219 patients receiving mechanical ventilation to assess ED ventilation practices. Median tidal volume was 7.6 mL/kg predicted body weight (PBW) (interquartile range, 6.9-8.9), with a range of 4.3 to 12.2 mL/kg PBW. Lung-protective ventilation was used in 122 patients (55.7%). The incidence of ARDS after admission from the ED was 14.7%, with a mean onset of 2.3 days. Progression to ARDS was associated with higher illness severity and intubation in the prehospital environment or transferring facility. Of the 15 patients with ARDS in the ED (6.8%), lung-protective ventilation was used in seven (46.7%). Patients who progressed to ARDS experienced greater duration in organ failure and ICU length of stay and higher mortality. Lung-protective ventilation is infrequent in patients receiving mechanical ventilation in the ED, regardless of ARDS status. Progression to ARDS is common after admission, occurs early, and worsens outcome. Patient- and treatment-related factors present in the ED are associated with ARDS. Given the limited treatment options for ARDS, and the early onset after admission from the ED, measures to prevent onset and to mitigate severity should be instituted in the ED. ClinicalTrials.gov; No.: NCT01628523; URL: www.clinicaltrials.gov.


Prenatal Phthalate Exposure and Childhood Growth and Blood Pressure: Evidence from the Spanish INMA-Sabadell Birth Cohort Study.
Human evidence on the effects of early life phthalate exposure on obesity and cardiovascular disease risks, reported by experimental studies, is limited to a few cross-sectional studies. We evaluated the associations between prenatal phthalate exposure and childhood growth and blood pressure in a Spanish birth cohort study. We assessed exposure using the average of two phthalate metabolite spot-urine concentrations collected from the mothers in the first and third pregnancy trimesters (creatinine-adjusted, n = 391). Study outcomes were the difference in age- and sex-specific z-scores for weight between birth and 6 months of age; and repeated age- and sex-specific z-scores for body mass index (BMI) at 1, 4, and 7 years; waist-to-height ratio at 4 and 7 years; and age- and height-specific z-scores for systolic and diastolic blood pressure at 4 and 7 years. The sum of five high-molecular-weight phthalate metabolites (ΣHMWPm) was associated with lower weight z-score difference between birth and 6 months (β per doubling of exposure = -0.41; 95% CI: -0.75, -0.06) and BMI z-scores at later ages in boys (β = -0.28; 95% CI: -0.60, 0.03) and with higher weight z-score difference (β = 0.24; 95% CI: -0.16, 0.65) and BMI z-scores in girls (β = 0.30; 95% CI: -0.04, 0.64) (p for sex interaction = 0.01 and 0.05, respectively). The sum of three low-molecular-weight phthalates (ΣLMWPm) was not significantly associated with any of the growth outcomes. ΣHMWPm and ΣLMWPm were associated with lower systolic blood pressure z-scores in girls but not in boys. This study suggests that prenatal phthalate exposure may be associated with postnatal growth and blood pressure in a sex-specific manner. Inconsistencies with previous cross-sectional findings highlight the necessity for evaluating phthalate health effects in prospective studies.


Tobacco control in schools of India: review from India Global School Personnel Survey 2006.
School personnel are role models for students, teachers of tobacco use prevention curricula, and key influencers for tobacco control policies in schools. With their daily interactions and strong influence on their students they represent an influential group for tobacco control. Data collected through the Global School Personnel Survey during 2006 in 180 school; of 6 regions of India have shown that a large proportion (29%) of school personnel used tobacco. The scarcity of tobacco free school policies and relevant teaching materials (non existent in 62% of schools surveyed) and lack of training among school personnel (84%) reported in this study indicate the extent of undermining the scope of prevention efforts in schools to reduce adolescent tobacco use prevalence in India. Majority of school personnel in India strongly agreed that they should receive specific training to help students avoid or stop using tobacco. Training of school personnel along with introduction of comprehensive school policies and its vigorous enforcement will help adolescent students and school personnel to adopt and maintain a tobacco free lifestyle.


Variational Bayesian image restoration based on a product of t-distributions image prior.
Image priors based on products have been recognized to offer many advantages because they allow simultaneous enforcement of multiple constraints. However, they are inconvenient for Bayesian inference because it is hard to find their normalization constant in closed form. In this paper, a new Bayesian algorithm is proposed for the image restoration problem that bypasses this difficulty. An image prior is defined by imposing Student-t densities on the outputs of local convolutional filters. A variational methodology, with a constrained expectation step, is used to infer the restored image. Numerical experiments are shown that compare this methodology to previous ones and demonstrate its advantages.


Cross talk between Paneth and tuft cells drives dysbiosis and inflammation in the gut mucosa.
Gut microbiota imbalance (dysbiosis) is increasingly associated with pathological conditions, both within and outside the gastrointestinal tract. Intestinal Paneth cells are considered to be guardians of the gut microbiota, but the events linking Paneth cell dysfunction with dysbiosis remain unclear. We report a three-step mechanism for dysbiosis initiation. Initial alterations in Paneth cells, as frequently observed in obese and inflammatorybowel diseases patients, cause a mild remodeling of microbiota, with amplification of succinate-producing species. SucnR1-dependent activation of epithelial tuft cells triggers a type 2 immune response that, in turn, aggravates the Paneth cell defaults, promoting dysbiosis and chronic inflammation. We thus reveal a function of tuft cells in promoting dysbiosis following Paneth cell deficiency and an unappreciated essential role of Paneth cells in maintaining a balanced microbiota to prevent inappropriate activation of tuft cells and deleterious dysbiosis. This succinate-tuft cell inflammation circuit may also contribute to the chronic dysbiosis observed in patients.


Interleukin-10 gene therapy-mediated amelioration of bacterial pneumonia.
Respiratory infection by Actinobacillus pleuropneumoniae causes a highly pathogenic necrotizing pleuropneumonia with severe edema, hemorrhage and fever. Acute infection is characterized by expression of inflammatory cytokines, including interleukin-1 (IL-1), IL-6 and IL-8. To determine if high level production of inflammatory cytokines contributed to disease pathogenesis, we investigated if inhibiting macrophage activation with adenovirus type 5-expressed IL-10 (Ad-5/IL-10) reduced the severity of acute disease. Porcine tracheal epithelial cells infected with Ad-5/IL-10 produced bioactive human IL-10. When pigs were intratracheally infected with A. pleuropneumoniae, pigs pretreated with Ad-5/IL-10 showed a significant reduction in the amount of lung damage when compared to adenovirus type 5-expressing beta-galactosidase (Ad-5/beta-Gal)-treated and untreated pigs. In addition, serum zinc levels were unchanged, the lung weight/body weight ratio (an indicator of vascular leakage) was significantly reduced, and lung pathology scores were reduced. Myeloperoxidase activity in lung lavage fluid samples, an indicator of neutrophil invasion, was decreased to levels similar to that seen in pigs not infected with A. pleuropneumoniae. Reduction in inflammatory cytokine levels in lung lavage fluid samples correlated with the clinical observations in that pigs pretreated with Ad-5/IL-10 showed a corresponding reduction of IL-1 and tumor necrosis factor (TNF) compared with untreated and Ad-5/beta-Gal-treated pigs. IL-6 levels were unaffected by pretreatment with Ad-5/IL-10, consistent with observations that IL-6 was not derived from alveolar macrophages. Since inflammatory cytokines are expressed at high levels in acute bacterial pleuropneumonia, these results indicate that macrophage activation, involving overproduction of IL-1 and TNF, is a prime factor in infection-related cases of massive lung injury.


Regional Variation in Muscle Echogenicity Is Related to Muscle Thickness in Young Children.
Quantitative ultrasound of muscle echogenicity may be influenced by the size of the muscle and, so, corrections may be required when comparing echogenicity between populations with different muscle size. This study examined the relationship between regional muscle echogenicity and geometry in young, typically developing children (n = 49, mean ± standard deviation [SD] age = 70.8 ± 30.0 mo). Three-dimensional ultrasound was used to measure mean echo intensity (EI) and echo variation (EV), together with muscle thickness, cross-sectional area (CSA) and width, over the entire muscle length for the medial gastrocnemius (MG), rectus femoris (RF) and tibialis anterior muscles. Pearson's correlation coefficient (r) was used to assess the strength of the relationship between echogenicity and geometry using all images taken over the entire muscle length. There were moderate-strong correlations (r = 0.67-0.90) between EI and EV and thickness for each muscle, with the strongest correlations evident for the MG and RF. EI and EV were moderately correlated with muscle CSA and weakly correlated with muscle width. Normalisation of echogenicity to muscle thickness may help delineate between regions of contractile and non-contractile tissue and provide a useful measure of muscle echogenicity when comparing muscles of different cross-sectional dimensions. We recommend that researchers consider controlling for muscle size-dependent effects on echogenicity, by normalising EI and EV to muscle thickness, or including the latter as a covariate when comparing between groups.


Glycation of apoprotein A-I is associated with coronary artery plaque progression in type 2 diabetic patients.
To investigate whether glycation level of apoprotein (apo)A-I is associated with coronary artery disease (CAD) and plaque progression in patients with type 2 diabetes. Among 375 consecutive type 2 diabetic patients undergoing quantitative coronary angiography (QCA) and intravascular ultrasound (IVUS), 82 patients with nonsignificant stenosis (luminal diameter narrowing <30% [group I]) and 190 patients with significant CAD (luminal diameter stenosis ≥70% [group II]) were included for analysis of apoA-I glycation level and serum activity of lecithin: cholesterol acyltransferase (LCAT). The control group had 136 healthy subjects. At the 1-year follow-up, angiography and IVUS were repeated mainly in group II patients for plaque progression assessment. Relative intensity of apoA-I glycation by densitometry was increased, and serum LCAT activity was decreased stepwise across groups control, I, and II. These two measurements were associated with the number of diseased coronary arteries and extent index in group II. During 1-year follow-up, QCA detected 45 patients with plaque progression in 159 subjects, and IVUS found 38 patients with plaque progression in 127 subjects. Baseline relative intensity of apoA-I glycation was significantly increased in patients with plaque progression compared with those without, with values associated with changes in QCA and IVUS measurements. Multivariable regression analysis revealed that baseline relative intensity of apoA-I glycation was an independent determinant of CAD and plaque progression in type 2 diabetic patients. ApoA-I glycation level is associated with the severity of CAD and coronary artery plaque progression in type 2 diabetic patients.


Clarithromycin and N-acetylcysteine co-spray-dried powders for pulmonary drug delivery: A focus on drug solubility.
Cystic fibrosis (CF) lungs are usually susceptible to Pseudomonas aeruginosa colonization and this bacterium is resistant to immune system clearance and drug control. Particularly, the biofilm mode of growth protects several microorganisms from host defenses and antibacterial drugs, mainly due to a delayed penetration of the drug through the biofilm matrix. Biofilm, together with lung mucus viscosity and tenacity, reduces, therefore, the effectiveness of conventional antibiotic therapy in CF. The aim of this research was to design and develop a stable, portable, easy to use dry powder inhaler (DPI) for CF patients, able to release directly to the lung an association of macrolide antibiotics (clarithromycin) and a mucolytic agent (N-Acetyl-Cysteine). Its effectiveness is based on the counteracting of the characteristics of P. aeruginosa infections in CF (lung bacterial adhesion to lung epithelium, biofilm formation and mucus viscosity) and the ability to let the antimicrobial drug exert their pharmacological action. A solution of these two drugs, without any excipients, was spray-dried to obtain respirable microparticles, characterized by aerodynamic diameters suitable for inhalation (<5.0μm). The morphology evaluation evidenced particles shape dependent on water content in the spray drying feeds, with wrinkled particles more evident with higher water content. Moreover, thanks to the presence of N-acetylcysteine which can interact with clarithromycin dimethyl-amino group, a consistent enhancement of drug solubility was obtained, compared to raw material and to the drug sprayed alone. The mucolytic agent added in the DPI may improve the macrolide diffusion into the mucus, enabling its action.


Physical activity as a risk factor for prostate cancer diagnosis: a prospective biopsy cohort analysis.
To assess the association between physical activity, evaluated by the Physical Activity Scale for the Elderly (PASE) questionnaire, and prostate cancer risk in a consecutive series of men undergoing prostate biopsy. From 2011 onwards, consecutive men undergoing 12-core prostate biopsy were enrolled into a prospective database. Indications for a prostatic biopsy were a prostate-specific antigen (PSA) value of ≥4 ng/mL and/or a positive digital rectal examination. Body mass index (BMI) and waist circumferences were measured before the biopsy. Fasting blood samples were collected before biopsy and tested for: total PSA, glucose, high-density lipoprotein cholesterol, and trygliceride levels. Blood pressure was recorded. Metabolic syndrome (MetS) was defined according to the Adult Treatment panel III. The PASE questionnaire was completed before the biopsy. In all, 286 patients were enrolled with a median (interquartile range, IQR) age and PSA level of 68 (62-74) years and 6.1 (5-8.8) ng/mL, respectively. The median (IQR) BMI was 26.4 (24.6-29) kg/m(2) and waist circumference was 102 (97-108) cm, with 75 patients (26%) presenting with MetS. In all, 106 patients (37%) had prostate cancer at biopsy. Patients with prostate cancer had higher PSA levels (median [IQR] 6.7 [5-10] vs 5.6 [4.8-8] ng/mL; P = 0.007) and lower LogPASE scores (median [IQR] 2.03 [1.82-2.18] vs 2.10 [1.92-2.29]; P = 0.005). On multivariate analysis, in addition to well-recognised risk factors such as age, PSA level and prostate volume, LogPASE score was an independent risk factor for prostate cancer diagnosis (odds ratio [OR] 0.146, 95% confidence interval [CI] 0.037-0.577; P = 0.006]. LogPASE score was also an independent predictor of high-grade cancer (OR 0.07, 95% CI 0.006-0.764; P = 0.029). In our single-centre study, increased physical activity, evaluated by the PASE questionnaire, is associated with a reduced risk of prostate cancer and of high-grade prostate cancer at biopsy. Further studies should clarify the molecular pathways behind this association.


Effects of mexiletine on electrical field stimulation-induced contractile responses in the ipsilateral and contralateral vasa deferentia after unilateral testicular torsion/detorsion.
To investigate testicular torsion-induced changes on the electrical field stimulation (EFS)-induced contractions in rabbit vasa deferentia and to evaluate the effect of mexiletine. 18 male New Zealand albino rabbits were used in this experiment. Rabbits were divided into three groups: (1) control group (n = 6); (2) torsion group (n = 6), and (3) mexiletine group (n = 6). In the control group, vasa deferentia on both sides were harvested. In the torsion and mexiletine groups, the left testes of the rabbits were subjected to 720 degrees of clockwise torsion for 2 h and then detorsion was performed. In the mexiletine group, 50 mg/kg i.p. mexiletine was administered 1 h before detorsion. Following 24 h of the torsion, vasa deferentia on both sides were harvested and 2-cm strips including both the prostatic and epididymal portions were prepared to record EFS-induced contractions. Testicular torsion caused a significant inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia. Mexiletine treatment did not affect these inhibitory responses. Torsion/detorsion of the spermatic cord did not alter exogenously applied noradrenaline-induced contractions in both vasa deferentia. However, KCl-induced contractions diminished significantly in ipsilateral vas deferens of the torsion group and mexiletine restored this inhibition. Unilateral testicular torsion/detorsion leads to inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia by causing a defect in presynaptic nerve transmission. However, mexiletine has no effect on this inhibition. Inhibition of the KCl-induced contractions in the ipsilateral vas deferens, which indicates postsynaptic tissue damage, is restored by administering mexiletine 1 h prior to detorsion.


Highly potent anti-CD20-RLI immunocytokine targeting established human B lymphoma in SCID mouse.
Rituximab (RTX), a chimeric IgG1 monoclonal antibody directed against the CD20 antigen, has revolutionized the treatment of B-cell malignancies. Nevertheless, the relapsed/refractory rates are still high. One strategy to increase the clinical effectiveness of RTX is based on antibody-cytokine fusion protein (immunocytokine; ICK) vectorizing together at the tumor site the antibody effector activities and the cytokine co-signal required for the generation of cytotoxic cellular immunity. Such ICKs linking various antibody formats to interleukin (IL)-2 are currently being investigated in clinical trials and have shown promising results in cancer therapies. IL-15, a structurally-related cytokine, is now considered as having a better potential than IL-2 in antitumor immunotherapeutic strategies. We have previously engineered the fusion protein RLI, linking a soluble form of human IL-15Rα-sushi+ domain to human IL-15. Compared with IL-15, RLI displayed better biological activities in vitro and higher antitumor effects in vivo in murine and human cancer models. In this study, we investigated the advantages of fusing RLI to RTX. Anti-CD20-RLI kept its binding capacity to CD20, CD16 and IL-15 receptor and therefore fully retained both antibody effector functions (ADCC and CDC), and the cytokine potential of RLI. In a severe combined immunodeficiency (SCID) mouse model of disseminated residual lymphoma, anti-CD20-RLI was found to induce long-term survival of 90% of mice up to at least 120 days whereas RLI and RTX, alone or in combination, just delayed the disease onset (100% of death at 28, 40 and 51 days respectively). These findings suggest that such ICK could improve the clinical efficacy of RTX, particularly in patients with refractory B-cell lymphoma.


Chemopreventive potential of Tribulus terrestris against 7,12- dimethylbenz (a) anthracene induced skin papillomagenesis in mice.
In the present investigation, the chemopreventive potential of aqueous extracts of the root and fruit of Tribulus terrestris (an Ayurvedic medicinal plant) on 7, 12 - dimethylbenz (a) anthracene (DMBA) induced papillomagenesis in male Swiss albino mice was studied. A significant reduction in tumor incidence, tumor burden and cumulative number of papillomas was observed, along with a significant increase in average latent period in mice treated orally with Tribulus terrestris suspension continuously at pre, peri and post-initiation stages of papillomagenesis as compared to the control group treated with DMBA and croton oil alone. Treatment with Tribulus terrestris suspension by oral gavage for 7 days resulted in a significant increase in the reduced glutathione content in the liver (P< 0.001 for both root and fruit extracts). Conversely, lipid peroxidation levels were significantly decreased (P< 0.001).


Neuroendocrine carcinoma of ovary: Hitherto rare entity in primary ovarian tumors.
Neuroendocrine neoplasms (NEN) of the female genital tract are extremely uncommon. These tumors can be broadly divided into well differentiated (carcinoid) and poorly differentiated NEN (small cell and large cell carcinomas). Occurrence of neuroendocrine carcinomas (NECs) in ovary has rarely been reported. These high-grade malignant tumors have a fulminant clinical course with a short period of survival, even when diagnosed at an early stage. We hereby report two cases of primary neuroendocrine carcinoma of the ovary.


Stent management of coil herniation in embolization of internal carotid aneurysms.
Coil herniation into the parent artery after detachment is an uncommon complication of embolization of the intracranial aneurysm. We report our experience with stent reconstruction of the lumen and flow of the internal carotid artery (ICA) after coil herniation during embolization for intracranial ICA aneurysms and the possible mechanisms of coil herniation. A series of 216 consecutive patients was treated by endovascular coil embolizations for intracranial aneurysms. Of these patients, there were 9 (4 men, 5 women; 32-68 years of age) complicating with coil herniation into the ICA and undergoing stent deployment to reconstruct the ICA lumen (n = 8) or both lumen and flow (n = 1). Wide-neck aneurysms were found in 8 and narrow-neck, in 1. Aneurysms were in the posterior communicating artery (n = 5) and the paraophthalmic (n = 3) and cavernous portions (n = 1) of the ICA. Self-expandable stents were deployed in the ICA in 6; balloon-mounted stents were selected in 3. The causes of coil herniation appeared to be coil instability after detachment (n = 6), excessive embolization (n = 1), microcatheter-related problems (n = 1), or being pushed by subsequent coil embolization (n = 1). Endovascular stent placement to reconstruct the lumen and/or flow of the ICA was technically successful in all 9 patients; 1 needed a second stent due to further coil migration. No significant procedure-related complications were found. Clinical follow-up was 8-35 months. Coil herniation occasionally occurs during endovascular embolization of ICA aneurysms because of coil instability after detachment, excessive embolization, microcatheter-related problems, or pushing by subsequent coil embolization. In this small series, stent placement was safe and effective in the reconstruction of the arterial lumen and/or restoration of flow past a herniated coil mass.


Disparities, variations, inequalities or inequities: whatever you call them, we need data to monitor them.
Health inequalities are a problem in high, middle and low income countries. Most are unfair ('inequities') and could be minimised but primarily through policies outside the health service.In the US, the Center for Diseases Control has used high quality, nationally-available data to monitor conditions and determinants of health among different groups (by sex, disability, race, ethnicity, and language) to motivate action to reduce inequalities. In the UK, the 10 top level 'health' indicators in London at the turn of the millennium included unemployment, education, housing quality, crime, air pollution, road travel injuries, as well as traditional health measures. Most of these affect mental and physical health through social determinants or adverse environmental exposures. Current inequalities monitoring in England includes a Local Basket of Inequalities Indicators focusing on a wide range of determinants of health as well as traditional health metrics.Israel, like the US, has above average socio-economic inequalities but has universal healthcare. Health inequalities in Israel occur within different Jewish groups and by income, education, ethnicity, and religion, with disadvantages often clustering. Current monitoring in Israel focuses on health outcomes and 'midstream' healthcare-related provision. I agree with Abu-Saad and her colleagues that including monitoring of social determinants of health is crucial to identify and tackle health inequalities in Israel.National, 'upstream', interventions are the most effective ways to reduce inequalities and improve the population's health. High-level political support is crucial for this. While a 'Health in all Policies' approach combined with political will to 'leave no one behind' can lead to great improvements, regular monitoring is essential, to: identify the inequities; plan appropriate and effective, targeted interventions; implement and evaluate them; and change them where needed. All of this requires adequate and timely data on health and its determinants, including information about undiagnosed and poorly controlled disease, obtained from the general population not just those attending for healthcare, analysed for each population sub-group at risk of experiencing inequalities.This is a commentary on https://doi.org/10.1186/s13584-018-0208-1.


[Effect of enterogastric circulation of diazepam on its CNS inhibitory action in mice].
The present study was designed to examine whether disturbance of the enterogastric circulation of diazepam would obviously affect its inhibitory action on CNS. Intragastric administration of acidic liquid (pH 1) elicited a marked increase in the amount of diazepam in the gastric juice of mice 1 h after i.v. diazepam, as measured by HPLC method. The effects of ig acidic charcoal (2 g.kg-1, pH 1), neutral charcoal and normal saline on diazepam-induced CNS depression were compared by studying the duration of hypnotic action, inhibition of spontaneous activity, dropping rate in rotating rod test and mortality after i.v. diazepam (10-40 mg.kg-1) in mice. The results showed that the animals receiving ig acidic charcoal recovered more rapidly from CNS depression and exhibited lower mortality than the animals in neutral charcoal group and normal saline group.


Identification of a novel repressor encoded by the putative gene ctf1 for cellulase biosynthesis in Trichoderma reesei through artificial zinc finger engineering.
Strains from Trichoderma reesei have been used for cellulase production with a long history. It has been well known that cellulase biosynthesis by the fungal species is controlled through regulators, and elucidation of their regulation network is of great importance for engineering T. reesei with robust cellulase production. However, progress in this regard is still very limited. In this study, T. reesei RUT-C30 was transformed with an artificial zinc finger protein (AZFP) library, and the mutant T. reesei M2 with improved cellulase production was screened. Compared to its parent strain, the filter paper activity and endo-β-glucanase activity in cellulases produced by T. reesei M2 increased 67.2% and 35.3%, respectively. Analysis by quantitative reverse transcription polymerase chain reaction indicated significant downregulation of the putative gene ctf1 in T. reesei M2, and its deletion mutants were thus developed for further studies. An increase of 36.9% in cellulase production was observed in the deletion mutants, but when ctf1 was constitutively overexpressed in T. reesei RUT-C30 under the control of the strong pdc1 promoter, cellulase production was substantially compromised. Comparative transcriptomic analysis revealed that the deletion of ctf1 upregulated transcription of gene encoding the regulator VIB1, but downregulated transcription of gene encoding another regulator RCE1, which consequently upregulated genes encoding the transcription factors XYR1 and ACE3 for the activation of genes encoding cellulolytic enzymes. As a result, ctf1 was characterized as a gene encoding a repressor for cellulase production in T. reesei RUT-C30, which is significant for further elucidating molecular mechanism underlying cellulase biosynthesis by the fungal species for rational design to develop robust strains for cellulase production. And in the meantime, AZFP transformation was validated to be an effective strategy for identifying functions of putative genes in the genome of T. reesei.


Perceptions of Clostridium difficile infections among infection control professionals in Taiwan.
High Clostridium difficile colonization and infection rates among hospitalized patients had been noted in Taiwan. Nevertheless, the cognition about clinical diagnosis and management of CDI among infection control professionals in Taiwan is not clear. A 24-item survey questionnaire about the diagnosis, therapy, or infection control policies toward CDI was distributed in the annual meeting of the Infectious Disease Society of Taiwan (IDST) in October 2015 and Infectious Control Society of Taiwan (ICST) in April 2016. Totally 441 individuals responded to the survey, and 280 (63.5%) participants would routinely monitor the prevalence of CDI and 347 (78.7%) reported the formulation of infection control policies of CDI in their hospital, including contact precaution (75.7%), wearing gloves (88.9%) or dressing (80.0%) at patient care, single room isolation (49.7%), preference of soap or disinfectant-based sanitizer (83.2%) and avoidance of alcohol-based sanitizer (63.3%), and environmental disinfection with 1000 ppm bleach (87.1%). For the timing of contact precaution discontinuation isolation for CDI patients, most (39.9%) participants suggested the time point of the absence of C. difficile toxin in feces. To treat mild CDI, most (61.9%) participants preferred oral metronidazole, and for severe CDI 26.1% would prescribe oral vancomycin as the drug of choice. There were substantial gaps in infection control polices and therapeutic choices for CDI between international guidelines and the perceptions of medical professionals in Taiwan. Professional education program and the setup of guideline for CDI should be considered in Taiwan.


Intact bacteria inhibit the induction of humoral immune responses to bacterial-derived and heterologous soluble T cell-dependent antigens.
During infections with extracellular bacteria, such as Streptococcus pneumoniae (Pn), the immune system likely encounters bacterial components in soluble form, as well as those associated with the intact bacterium. The potential cross-regulatory effects on humoral immunity in response to these two forms of Ag are unknown. We thus investigated the immunologic consequences of coimmunization with intact Pn and soluble conjugates of Pn-derived proteins and polysaccharides (PS) as a model. Coimmunization of mice with Pn and conjugate resulted in marked inhibition of conjugate-induced PS-specific memory, as well as primary and memory anti-protein Ig responses. Inhibition occurred with unencapsulated Pn, encapsulated Pn expressing different capsular types of PS than that present in the conjugate, and with conjugate containing protein not expressed by Pn, but not with 1-microm latex beads in adjuvant. Inhibition was long-lasting and occurred only during the early phase of the immune response, but it was not associated with tolerance. Pn inhibited the trafficking of conjugate from the splenic marginal zone to the B cell follicle and T cell area, strongly suggesting a potential mechanism for inhibition. These data suggest that during infection, bacterial-associated Ags are the preferential immunogen for antibacterial Ig responses.


Cryptic ecological diversification of a planktonic estuarine copepod, Acartia tonsa.
The recent discovery of cryptic species in marine holoplankton, organisms that 'drift' in oceanic currents throughout their life cycle, contrasts with their potential for long-distance passive dispersal and presumably high gene flow. These observations suggest that holoplankton species are adapting to surprisingly small-scale oceanographic features and imply either limited dispersal or strong selection gradients. Acartia tonsa is a widespread and numerically dominant estuarine copepod containing deep mitochondrial lineages within and among populations along the northwestern Atlantic coast. In this study, we intensively investigated A. tonsa populations in Chesapeake Bay with the goals of testing species status for the deep lineages and testing for their association with environmental features over space and time. Phylogenetic analyses of DNA sequences from mitochondrial cytochrome c oxidase I (mtCOI) and the nuclear ribosomal internal transcribed spacer (nITS) resolved two concordant monophyletic clades. Deep divergence between the two clades (13.7% uncorrected sequence divergence for mtCOI and 32.2% for nITS) and genealogical concordance within sympatric populations strongly suggest that the two clades represent reproductively isolated cryptic species. Based on restriction fragment length polymorphisms of mtCOI, representatives from the two clades were found consistently associated with contrasting salinity regimes (oligohaline vs. meso-polyhaline) with an overlap between 2 and 12 PSU in samples from 1995 to 2005. Finding these patterns in one of the best-known estuarine copepods reinforces the conclusion that marine biodiversity is underestimated, not only in terms of species numbers, but also with respect to niche partitioning and the potential importance of ecological divergence in marine holoplankton.


Genetic analysis of quantitative phenotypes in AD and MCI: imaging, cognition and biomarkers.
The Genetics Core of the Alzheimer's Disease Neuroimaging Initiative (ADNI), formally established in 2009, aims to provide resources and facilitate research related to genetic predictors of multidimensional Alzheimer's disease (AD)-related phenotypes. Here, we provide a systematic review of genetic studies published between 2009 and 2012 where either ADNI APOE genotype or genome-wide association study (GWAS) data were used. We review and synthesize ADNI genetic associations with disease status or quantitative disease endophenotypes including structural and functional neuroimaging, fluid biomarker assays, and cognitive performance. We also discuss the diverse analytical strategies used in these studies, including univariate and multivariate analysis, meta-analysis, pathway analysis, and interaction and network analysis. Finally, we perform pathway and network enrichment analyses of these ADNI genetic associations to highlight key mechanisms that may drive disease onset and trajectory. Major ADNI findings included all the top 10 AD genes and several of these (e.g., APOE, BIN1, CLU, CR1, and PICALM) were corroborated by ADNI imaging, fluid and cognitive phenotypes. ADNI imaging genetics studies discovered novel findings (e.g., FRMD6) that were later replicated on different data sets. Several other genes (e.g., APOC1, FTO, GRIN2B, MAGI2, and TOMM40) were associated with multiple ADNI phenotypes, warranting further investigation on other data sets. The broad availability and wide scope of ADNI genetic and phenotypic data has advanced our understanding of the genetic basis of AD and has nominated novel targets for future studies employing next-generation sequencing and convergent multi-omics approaches, and for clinical drug and biomarker development.


Vendor Independent Coronary Calcium Scoring Improves Individual Risk Assessment: MESA (Multi-Ethnic Study of Atherosclerosis).
Substantial variation in Agatston scores (AS) acquired with different computed tomography (CT) scanners may influence patient risk classification. This study sought to develop a calibration tool for state-of-the-art CT systems resulting in vendor-neutral AS (vnAS), and to assess the impact of vnAS on coronary heart disease (CHD) event prediction. The vnAS calibration tool was derived by imaging 2 anthropomorphic calcium containing phantoms on 7 different CT and 1 electron beam tomography system, which was used as the reference system. The effect of vnAS on CHD event prediction was analyzed with data from 3,181 participants from MESA (Multi-Ethnic Study on Atherosclerosis). Chi-square analysis was used to compare CHD event rates between low (vnAS <100) and high calcium groups (vnAS ≥100). Multivariable Cox proportional hazard regression models were used to assess the incremental value of vnAS. For all CT systems, a strong correlation with electron beam tomography-AS was found (R2 >0.932). Of the MESA participants originally in the low calcium group (n = 781), 85 (11%) participants were reclassified to a higher risk category based on the recalculated vnAS. For reclassified participants, the CHD event rate of 15% was significantly higher compared with participants in the low calcium group (7%; P = 0.008) with a CHD HR of 3.39 (95% CI: 1.82-6.35; P = 0.001). The authors developed a calibration tool that enables calculation of a vnAS. MESA participants who were reclassified to a higher calcium category by means of the vnAS experienced more CHD events, indicating improved risk categorization.


Patterned crystal growth and heat wave generation in hydrogels.
The crystallization of metastable liquid phase change materials releases stored energy as latent heat upon nucleation and may therefore provide a triggerable means of activating downstream processes that respond to changes in temperature. In this work, we describe a strategy for controlling the fast, exothermic crystallization of sodium acetate from a metastable aqueous solution into trihydrate crystals within a polyacrylamide hydrogel whose polymerization state has been patterned using photomasks. A comprehensive experimental study of crystal shapes, crystal growth front velocities and evolving thermal profiles showed that rapid growth of long needle-like crystals through unpolymerized solutions produced peak temperatures of up to 45˚C, while slower-crystallizing polymerized solutions produced polycrystalline composites and peaked at 30˚C due to lower rates of heat release relative to dissipation in these regions. This temperature difference in the propagating heat waves, which we describe using a proposed analytical model, enables the use of this strategy to selectively activate thermoresponsive processes in predefined areas.


Tourette syndrome: evolving concepts.
Tourette syndrome is a common childhood-onset neurobehavioral disorder characterized by multiple motor and phonic tics affecting boys more frequently than girls. Premonitory sensory urges prior to tic execution are common, and this phenomenon helps to distinguish tics from other hyperkinetic movement disorders. Tourette syndrome is commonly associated with attention deficit hyperactivity disorder, obsessive-compulsive disorder, learning difficulties, and impulse control disorder. The pathophysiology of this complex disorder is not well understood. Involvement of basal ganglia-related circuits and dopaminergic system has been suggested by various imaging and postmortem studies. Although it is considered a genetic disorder, possibly modified by environmental factors, an intense search has thus far failed to find causative genes. Symptomatic treatment of tics chiefly utilizes various alpha adrenergic agonists, antidopaminergic drugs, topiramate, botulinum toxin, and deep brain stimulation. Habit reversal therapy and other behavioral approaches may be a reasonable option for some cases. Improved understanding of Tourette syndrome should lead to better symptomatic and more effective pathogenesis-targeted therapies.


Association of total plasma homocysteine with methylenetetrahydrofolate reductase genotypes 677C>T, 1298A>C, and 1793G>A and the corresponding haplotypes in Swedish children and adolescents.
We studied 692 Swedish children and adolescents (aged 9-10 or 15-16 years, respectively), in order to evaluate the effect of the methylenetetrahydrofolate reductase (MTHFR) 677C>T, 1298A>C, and 1793G>A polymorphisms on total plasma homocysteine concentrations (tHcy). Genotyping was performed with Pyrosequencing technology. The MTHFR 677C>T polymorphism was associated with increased tHcy concentrations in both the children and the adolescents (P<0.001 for both age groups) in both genders. The effect of MTHFR 1298A>C was studied separately in subjects with the 677CC and 677CT genotypes, and the 1298C allele was found to be associated with higher tHcy levels both when children were stratified according to 677C>T genotypes, and when using haplotype analyses and diplotype reconstructions. The 1793A allele was in complete linkage disequilibrium with the 1298C allele. It was still possible to show that the 1793A allele was associated with lower tHcy levels, statistically significant in the adolescents. In conclusion, a haplotype-based approach was slightly superior in explaining the genetic interaction on tHcy plasma levels in children and adolescents than a simple genotype based approach (R2 adj 0.44 vs. 0.40). The major genetic impact on tHcy concentrations is attributable to the MTHFR 677C>T polymorphism. The common 1298A>C polymorphism had a minor elevating effect on tHcy, whereas the 1793G>A polymorphism had a lowering effect on tHcy.


Characteristics of Men With Peyronie's Disease and Collagenase Clostridium Histolyticum Treatment Failure: Predictors of Surgical Intervention and Outcomes.
A subset of patients with Peyronie's disease (PD) treated with collagenase clostridium histolyticum (CCH) experience persistent bother and some require surgery. We characterize patients experiencing persistent bother after CCH treatment and identify associations and predictors of surgical intervention/outcomes. We retrospectively identified patients with PD from October 2014 to October 2019 and identified those presenting with persistent bother after CCH treatment by other urologists. Intracavernosal injection and penile Doppler ultrasonography were performed, and subsequent interventions/outcomes were recorded. Baseline characteristics were compared with Student t-test and chi-square test. Predictors of surgical intervention and complications were assessed using multivariable logistic regression. The primary outcome was surgery after CCH treatment. Secondary outcomes included worsened erectile function, sensory deficits, and penile length change. Of 573, 67 (11.7%) patients with PD had undergone prior CCH treatment with median 6 injections (range 2-24). Mean post-CCH PD Questionnaire bother score was 10.1 (SD: 3.1), and total International Index of Erectile Function-5 was 15.3 (SD: 8.7). Mean PD duration was 27.8 (SD: 35.7) months, with a mean composite curvature (MCC) of 69.0° (SD: 33.8) measured after injection. Of 67, 44 (65.7%) patients had MCC >60°. Of 67, 52 (77.6%) patients had indent, narrowing, or hourglass and 26 (38.8%) had hinge effect (buckling of the erect penis with axial pressure) on examination. Calcification was identified in 26 of 67 (38.8%) patients, with grade 3 calcifications comprising 6 of 23 (26.1%) patients. Of 67, 33 (49.3%) patients underwent surgery, with 20 (60.1%) undergoing partial plaque excision and grafting with/without tunica albuginea plication, 6 (18.2%) undergoing tunica albuginea plication alone, and 7 (21.2%) undergoing penile prosthesis with plaque incision and grafting. Surgical patients had greater mean curvature (82.6 vs 55.4, P = .001) and were more likely to have hinge (54.5% vs 20.6%, P = .005). On multivariable analysis, MCC ≥60° predicted patient's decision for surgery (odds ratio: 2.99, P < .01, 95% confidence interval: 1.62-4.35). There were no associations between surgical complications and number of injections or CCH-associated adverse events. Patients presenting with persistent bother after CCH treatment often have narrowing and calcifications (despite calcifications being a contraindication to CCH treatment), and those who have hinge or severe curvature are more likely to undergo surgery with low rates of complications. This study's generalizability is limited by selection bias, but useful data are provided for patient counseling. Patients with persistent bother after CCH treatment had high rates of indentation/narrowing, plaque calcifications, and MCC >60° at completion of CCH treatment. Surgical intervention is more common with hinge and is safe and feasible in these patients, with low rates of complications. These findings suggest possible negative prognostic factors for CCH treatment, which merit further investigation. Bajic P, Wiggins AB, Ziegelmann MJ, et al. Characteristics of Men With Peyronie's Disease and Collagenase Clostridium Histolyticum Treatment Failure: Predictors of Surgical Intervention and Outcomes. J Sex Med 2020;17:1005-1011.


Analysis of the Possible Persistent Genotoxic Damage in Workers Linked to the Ardystil Syndrome.
A combination of several factors including a change in the paint application system; a lack of proper hygiene; and inadequate safety measures caused a severe health impact in the workers of some textile painting factories. This outbreak, mainly characterized by respiratory disorders, caused the death of six people and it has been classified as Ardystil syndrome. Fifty-two workers involved in the outbreak and 48 healthy subjects not known to have exposed to the potentially mutagenic agents participated in the study. The program evaluated possible genotoxic damage through the sister chromatid exchange (SCE) cytogenetic biomarker assay. We determined the frequency of SCE, high-frequency cells (HFCs), and a ratio, which can be considered as a new parameter, allowing for the study of the SCE distribution pattern among the chromosomes. There was no statistically significant difference in the SCE frequency and in the mean number of HFCs between the control and the Ardystil-affected groups. However, smoking increased the incidence of all parameters studied in both the case and control groups. This study shows that workers involved in the Ardystil syndrome did not suffer genotoxic damage as measured by SCE and HFCs when compared with the control group.


S-acylation of SOD1, CCS, and a stable SOD1-CCS heterodimer in human spinal cords from ALS and non-ALS subjects.
Previously, we found that human Cu, Zn-superoxide dismutase (SOD1) is S-acylated (palmitoylated) in vitro and in amyotrophic lateral sclerosis (ALS) mouse models, and that S-acylation increased for ALS-causing SOD1 mutants relative to wild type. Here, we use the acyl resin-assisted capture (acyl-RAC) assay to demonstrate S-acylation of SOD1 in human post-mortem spinal cord homogenates from ALS and non-ALS subjects. Acyl-RAC further revealed that endogenous copper chaperone for SOD1 (CCS) is S-acylated in both human and mouse spinal cords, and in vitro in HEK293 cells. SOD1 and CCS formed a highly stable heterodimer in human spinal cord homogenates that was resistant to dissociation by boiling, denaturants, or reducing agents and was not observed in vitro unless both SOD1 and CCS were overexpressed. Cysteine mutations that attenuate SOD1 maturation prevented the SOD1-CCS heterodimer formation. The degree of S-acylation was highest for SOD1-CCS heterodimers, intermediate for CCS monomers, and lowest for SOD1 monomers. Given that S-acylation facilitates anchoring of soluble proteins to cell membranes, our findings suggest that S-acylation and membrane localization may play an important role in CCS-mediated SOD1 maturation. Furthermore, the highly stable S-acylated SOD1-CCS heterodimer may serve as a long-lived maturation intermediate in human spinal cord.


Caprine arthritis-encephalitis: clinical features and presence of antibody in selected goat populations.
Features of caprine arthritis-encephalitis, a retrovirus disease of domestic goats, were studied in 60 goats over a 10-year period. The rate of progression and the severity of the disease process were highly variable within and among animals, but the most salient features were chronically swollen joints and bursae, lameness, weight loss, poor coat, mineralization of soft tissue, and death. Of 1,160 goat serum samples from 24 states tested by the immunodiffusion technique, 81% were positive for antibody to caprine arthritis-encephalitis virus antigens.


The Caenorhabditis elegans Y87G2A.14 Nudix hydrolase is a peroxisomal coenzyme A diphosphatase.
The number of Nudix hydrolase family members varies widely among different organisms. In order to understand the reasons for the particular spectrum possessed by a given organism, the substrate specificity and function of different family members must be established. The Y87G2A.14 Nudix hydrolase gene product of Caenorhabditis elegans has been expressed as a thioredoxin fusion protein in Escherichia coli and shown to be a CoA diphosphatase with catalytic activity towards CoA and its derivatives. The products of CoA hydrolysis were 3',5'-ADP and 4'-phosphopantetheine with Km and kcat values of 220 microM and 13.8 s(-1) respectively. CoA esters yielded 3',5'-ADP and the corresponding acyl-phosphopantetheine. Activity was optimal at pH 9.5 with 5 mM Mg2+ and fluoride was inhibitory with a Ki of 3 microM. The Y87G2A.14 gene product has a potential C-terminal tripeptide PTS1 peroxisomal targeting signal - SKI. By fusing a Y87G2A.14 cDNA to the C-terminus of yeast-enhanced green fluorescent protein, the enzyme appeared to be targeted to peroxisomes by the SKI signal when transfected into yeast cells. Deletion of SKI abolished specific targeting. The presence of related sequences with potential PTS1 or PTS2 peroxisomal targeting signals in other organisms suggests a conserved peroxisomal function for the CoA diphosphatase members of this group of Nudix hydrolases.


The significance of cysteine synthesis for acclimation to high light conditions.
Situations of excess light intensity are known to result in the emergence of reactive oxygen species that originate from the electron transport chain in chloroplasts. The redox state of glutathione and its biosynthesis contribute importantly to the plant's response to this stress. In this study we analyzed the significance of cysteine synthesis for long-term acclimation to high light conditions in Arabidopsis thaliana. Emphasis was put on the rate-limiting step of cysteine synthesis, the formation of the precursor O-acetylserine (OAS) that is catalyzed by serine acetyltransferase (SERAT). Wild type Arabidopsis plants responded to the high light condition (800 μmol m(-2) s(-1) for 10 days) with synthesis of photo-protective anthocyanins, induction of total SERAT activity and elevated glutathione levels when compared to the control condition (100 μmol m(-2) s(-1)). The role of cysteine synthesis in chloroplasts was probed in mutant plants lacking the chloroplast isoform SERAT2;1 (serat2;1) and two knock-out alleles of CYP20-3, a positive interactor of SERAT in the chloroplast. Acclimation to high light resulted in a smaller growth enhancement than wild type in the serat2;1 and cyp20-3 mutants, less induction of total SERAT activity and OAS levels but similar cysteine and glutathione concentrations. Expression analysis revealed no increase in mRNA of the chloroplast SERAT2;1 encoding SERAT2;1 gene but up to 4.4-fold elevated SERAT2;2 mRNA levels for the mitochondrial SERAT isoform. Thus, lack of chloroplast SERAT2;1 activity or its activation by CYP20-3 prevents the full growth response to high light conditions, but the enhanced demand for glutathione is likely mediated by synthesis of OAS in the mitochondria. In conclusion, cysteine synthesis in the chloroplast is important for performance but is dispensable for survival under long-term exposure to high light and can be partially complemented by cysteine synthesis in mitochondria.