[Direct effect of met-enkephalin on the synthesis and secretion of corticosterone by the adrenal glands of the rat].
ACTH (1 unit per 1 rat) induced a 6-fold increase of the corticosterone synthesis in adrenals and a 2-fold increase of its secretion into the blood. Met-enkephalin (15 micrograms/kg) also increase the corticosterone secretion, the level of the hormone in the blood, however, becoming even lower. Combined action of these two substances involved a greater increase of the hormone concentration in the gland than in case of each of them separately. The hormone content in the blood remained low in this case, too. In perfusion of an isolated adrenal gland with met-enkephalin (15 micrograms/ml), no increase in the corticosterone concentration was observed either in the perfusate samples. The data obtained suggest that met-enkephalin activates synthesis of corticosteroids, on the one hand, and inhibits their secretion into the blood, on the other hand, whereas ACTH activates both these processes.


Protection against cisplatin ototoxicity in a Sprague-Dawley rat animal model.
Cisplatin (CDDP) is an anti-neoplastic drug extensively used in cases of head and neck cancer. Cisplatin induces numerous untoward side-effects including ototoxicity. In this study, cisplatin ototoxicity in Sprague-Dawley rat animal model has been evaluated and the oto-protection provided by the systemic administration of the antioxidant drug D-methionine has been tested. A total of 12 Sprague-Dawley rats were used: 8 were treated intra-peritoneally with D-methionine (300 mg/kg) and cisplatin (16 mg/kg, slow 30 min-infusion), 4 only with cisplatin. The hearing threshold of the animals was evaluated by electrophysiological procedures as Otoacoustic Emissions and Auditory Brainstem Responses. The effects of protection were evaluated after 72 hours. The data from the Otoacoustic Emissions (in the 4.0-12 kHz band) and Auditory Brainstem Responses recordings suggested that D-methionine can partially protect from Cisplatin ototoxicity.


Mechanisms of impaired beta-adrenoceptor-induced airway relaxation by interleukin-1beta in vivo in the rat.
We studied the in vivo mechanism of beta-adrenergic receptor (beta-AR) hyporesponsiveness induced by intratracheal instillation of interleukin-1beta (IL-1beta, 500 U) in Brown-Norway rats. Tracheal and bronchial smooth muscle responses were measured under isometric conditions ex vivo. Contractile responses to electrical field stimulation and to carbachol were not altered, but maximal relaxation induced by isoproterenol (10(-6)-10(-5) M) was significantly reduced 24 h after IL-1beta treatment in tracheal tissues and to a lesser extent, in the main bronchi. Radioligand binding using [125I]iodocyanopindolol revealed a 32+/-7% reduction in beta-ARs in lung tissues from IL-1beta-treated rats, without any significant changes in beta2-AR mRNA level measured by Northern blot analysis. Autoradiographic studies also showed significant reduction in beta2-AR in the airways. Isoproterenol-stimulated cyclic AMP accumulation was reduced by IL-1beta at 24 h in trachea and lung tissues. Pertussis toxin reversed this hyporesponsiveness to isoproterenol but not to forskolin in lung tissues. Western blot analysis revealed an IL-1beta-induced increase in Gi(alpha) protein expression. Thus, IL-1beta induces an attenuation of beta-AR-induced airway relaxation through mechanisms involving a reduction in beta-ARs, an increase in Gi(alpha) subunit, and a defect in adenylyl cyclase activity.


Synthesis of biomolecule-modified mesoporous silica nanoparticles for targeted hydrophobic drug delivery to cancer cells.
Synthetic methodologies integrating hydrophobic drug delivery and biomolecular targeting with mesoporous silica nanoparticles are described. Transferrin and cyclic-RGD peptides are covalently attached to the nanoparticles utilizing different techniques and provide selectivity between primary and metastatic cancer cells. The increase in cellular uptake of the targeted particles is examined using fluorescence microscopy and flow cytometry. Transferrin-modified silica nanoparticles display enhancement in particle uptake by Panc-1 cancer cells over that of normal HFF cells. The endocytotic pathway for these particles is further investigated through plasmid transfection of the transferrin receptor into the normal HFF cell line, which results in an increase in particle endocytosis as compared to unmodified HFF cells. By designing and attaching a synthetic cyclic-RGD, selectivity between primary cancer cells (BT-549) and metastatic cancer cells (MDA-MB 435) is achieved with enhanced particle uptake by the metastatic cancer cell line. Incorporation of the hydrophobic drug Camptothecin into these two types of biomolecular-targeted nanoparticles causes an increase in mortality of the targeted cancer cells compared to that caused by both the free drug and nontargeted particles. These results demonstrate successful biomolecular-targeted hydrophobic drug delivery carriers that selectively target specific cancer cells and result in enhanced drug delivery and cell mortality.


[The role of dopamine receptors in controlling mouse aggressivity: the genotype dependence].
Significant genotypic differences in shock-induced aggression were found in mice of eight inbred strains. Aggression was evaluated in test with the action of low electric current through the cage floor. Low aggressive strains C3H/He, DD, BALB/c, AKR and highly aggressive strains CBA, DBA/2. CC57Br were singled out by the number of aggressive attacks. Selective stimulation of dopamine D2 receptors by bromocriptine considerably increased the shock-induced aggressiveness in mice of low-aggressive strains. Blockade of D2-receptors by the injection of antagonist sulpiride decreased or prevented the manifestation of aggression in highly-aggressive mice. At the same time selective agonist of dopamine (D1) receptors SKF 38393 and administration of selective antagonist of D1-receptors SCH 23390 did not influence significantly shock-induced aggression. Thus, shock-induced aggression, depends on the animal genotype and activation of D2-receptors.


Biochemical Characterization of QPX7728, a New Ultrabroad-Spectrum Beta-Lactamase Inhibitor of Serine and Metallo-Beta-Lactamases.
QPX7728 is a new ultrabroad-spectrum inhibitor of serine and metallo-beta-lactamases (MBLs) from a class of cyclic boronates that gave rise to vaborbactam. The spectrum and mechanism of beta-lactamase inhibition by QPX7728 were assessed using purified enzymes from all molecular classes. QPX7728 inhibits class A extended-spectrum beta-lactamases (ESBLs) (50% inhibitory concentration [IC50] range, 1 to 3 nM) and carbapenemases such as KPC (IC50, 2.9 ± 0.4 nM) as well as class C P99 (IC50 of 22 ± 8 nM) with a potency that is comparable to or higher than recently FDA-approved beta-lactamase inhibitors (BLIs) avibactam, relebactam, and vaborbactam. Unlike those other BLIs, QPX7728 is also a potent inhibitor of class D carbapenemases such as OXA-48 from Enterobacteriaceae and OXA enzymes from Acinetobacter baumannii (OXA-23/24/58, IC50 range, 1 to 2 nM) as well as MBLs such as NDM-1 (IC50, 55 ± 25 nM), VIM-1 (IC50, 14 ± 4 nM), and IMP-1 (IC50, 610 ± 70 nM). Inhibition of serine enzymes by QPX7728 is associated with progressive inactivation with a high-efficiency k2/K ranging from 6.3 × 104 (for P99) to 9.9 × 105 M-1 s-1 (for OXA-23). This inhibition is reversible with variable stability of the QPX7728-beta-lactamase complexes with target residence time ranging from minutes to several hours: 5 to 20 min for OXA carbapenemases from A. baumannii, ∼50 min for OXA-48, and 2 to 3 h for KPC and CTX-M-15. QPX7728 inhibited all tested serine enzymes at a 1:1 molar ratio. Metallo-beta-lactamases NDM, VIM, and IMP were inhibited by a competitive mechanism with fast-on-fast-off kinetics, with Ki s of 7.5 ± 2.1 nM, 32 ± 14 nM, and 240 ± 30 nM for VIM-1, NDM-1, and IMP-1, respectively. QPX7728's ultrabroad spectrum of BLI inhibition combined with its high potency enables combinations with multiple different beta-lactam antibiotics.


[Effect of M-cholinomimetics on nociceptive stimulation in the spinal cord].
Two series of experiments were performed to study the effects of oxotremorine (5, 10 and 20 micrograms/kg), arecoline (0.25, 0.5 and 1.0 mg/kg), pilocarpine (5, 10 and 20 mg/kg) and aceclidine (0.25, 0.5 and 1.0 mg/kg) injected intravenously on nociceptive transmission in the spinal cord of spinal rats. It was established that the m-cholinomimetics inhibit in a dose-dependent manner the nociceptive reflex induced by intraarterial injection of bradykinin. When given in the same doses the drugs reduce the bradykinin-induced bioelectrical activity in the afferent tracts of the spinal cord. Atropine (1 mg/kg i. v.) abolishes the antinociceptive action of the cholinomimetics.


[The age-specific changes in the wall of the arterial circle of Willis].
The objective of the present study was to create the database of morphometric and morphological characteristics of the wall of the vessels of the Willis arterial circle at the site of their bifurcation throughout the period of postnatal ontogenesis in man to be used for the postmortem determination of human age. Preparations of vascular cross sections were used to measure the intima thickness (intimal pads) in the region of lateral bifurcation angles on the terminal branch of the internal carotid and basilar arteries obtained from 130 corpses of the subjects aged between 0 and 75 years. The histological preparations were stained with hematoxylin/eosin and van Gieson's pyrofuchsin or by the Unnar-Tenzer method. Morphometry was performed using a Bioscan image analyser and the Scion Image v.402 software. The Microsoft Excel 2003 package was used for the primary treatment of the data obtained. They were used to draw the table of intimal pads developing throughout the period of postnatal ontogenesis in man that allows for the postmortem determination of approximate human age. The study showed that the thickening of intimal pads and concomitant thinning of the underlying muscular layer during the postnatal ontogenesis should be regarded as being normal age-related changes.


Transmembrane Na+ and Ca2+ electrochemical gradients in cardiac muscle and their relationship to force development.
Na+- and CA2+-sensitive microelectrodes were used to measure intracellular Na+ and Ca2+ activities (alpha iCa) of sheep ventricular muscle and Purkinje strands to study the interrelationship between Na+ and Ca2+ electrochemical gradients (delta muNa and delta muCa) under various conditions. In ventricular muscle, alpha iNa was 6.4 +/- 1.2 mM and alpha iCa was 87 +/- 20 nM ([Ca/+] = 272 nM). A graded decrease of external Na+ activity (alpha oNa) resulted in decrease of alpha iNa, and increase of alpha iCa. There was increase of twitch tension in low-alpha oNa solutions, and occasional increase of resting tension in 40% alpha oNa. Increase of external Ca2+ (alpha oCa) resulted in increase of alpha iCa and decrease of alpha iNa. Decrease of alpha oCa resulted in decrease of alpha iCa and increase of alpha iNa. The apparent resting Na-Ca energy ratio (delta muCa/delta muNa) was between 2.43 and 2.63. When the membrane potential (Vm) was depolarized by 50 mM K+ in ventricular muscle, Vm depolarized by 50 mV, alpha iNa decreased, and alpha iCa increased, with the development of a contracture. The apparent energy coupling ratio did not change with depolarization. 5 x 10(-6) M ouabain induced a large increase in alpha iNa ad alpha iCa, accompanied by an increase in twitch and resting tension. Under the conditions we have studied, delta muNa and delta muCa appeared to be coupled and n was nearly constant at 2.5, as would be expected if the Na-Ca exchange system was able to set the steady level of alpha iCa. Tension threshold was about 230 nM alpha iCa. The magnitude of twitch tension was directly related to alpha iCa.


Selenium accumulation protects plants from herbivory by Orthoptera via toxicity and deterrence.
To investigate whether selenium (Se) accumulation in plants provides a chemical defense against generalist insect herbivores, the feeding preference and performance of a mix of orthopteran species were investigated. The selenium hyperaccumulator Stanleya pinnata and accumulator Brassica juncea were used in herbivory studies in the laboratory, and S. pinnata was also used in a manipulative field experiment. In laboratory studies, both crickets and grasshoppers avoided plants pretreated with selenate, while those given no choice died after eating leaves with elevated Se (447 +/- 68 and 230 +/- 68 microg Se g(-1) DW, respectively). B. juncea has previously been shown to accumulate selenate, while S. pinnata hyperaccumulates methyl-selenocysteine. Thus, these findings demonstrate that both inorganic and organic forms of selenium protect plants from herbivory. Grasshoppers fed S. pinnata contained methylselenocysteine in their midgut and absorbed this form into surrounding tissues. In a manipulative field experiment, methylselenocysteine protected S. pinnata from invertebrate herbivory and increased its long-term survival rate over an entire growth season. * In native habitats of selenium hyperaccumulators, orthopterans represent a major group of insect herbivores. Protection offered by organic selenium accumulation against these herbivores may have promoted the evolution of selenium hyperaccumulation in plants.


Evaluation of elements in hair samples of children with developmental language disorder (DLD).
Recent studies have highlighted a role for trace elements and toxic metals across neurodevelopmental disorders, including developmental stuttering, Autistic Spectrum Disorders (ASD), and Attention Deficit/Hyperactivity Disorder (ADHD). However, these environmental influences have yet to be explored in relation to Developmental Language Disorder (DLD). Elemental hair composition of seven elements; zinc (64Zn), magnesium (26Mg), iron (57Fe), potassium (39K), aluminum (27Al), lead (208Pb), and barium (138Ba) were analyzed in hair samples from 35 children affected by DLD and 35 controls with typical language development (TLD) using both inductive coupled plasma optical emission spectroscopy (ICP-OES) and inductive coupled plasma mass spectroscopy (ICP-MS). The concentration of 64Zn was significantly lower in the hair of DLD group compared to the TLD control group. All other elements showed similar levels between cases and controls. This pilot study demonstrates the utility of trace elements and toxic metals screening in relation to language disorders and the use of hair samples in such investigations. The finding that zinc levels differed between cases and controls could represent a clinically relevant result and should be replicated in larger sample size across time. A wider battery of related elements will help to better understand the role of trace elements and toxic metals in DLD.


Preoperative transfusion versus no transfusion policy in sickle cell disease patients: a randomized trial.
Many children with sickle cell disease (SCD) indicated for adenotonsillectomy receive pre-operative transfusion therapy, either simple or exchange transfusion, in order to reduce surgical and sickle cell disease-related complications. This is a prospective randomized controlled clinical trial aiming to compare between preoperative simple transfusion and no transfusion in pediatric patients with sickle SCD admitted in Sultan Qaboos University Hospital, Muscat, Oman for adenotonsillectomy during the period from January 2014 through June 2018. They were randomly assigned into two arms (simple transfusion and no transfusion). Postoperative SCD-related complications have been encountered in 6 out of 138 patients (4.3%). There was no statistically significant difference between the two studied groups as regards the development of surgical or SCD-related complications (p = 0.6 and 0.8 respectively). The length of postoperative hospital stay was comparable in the two groups. (p = 0.607). SCD-related complications occurred exclusively in cases with homozygous sickle anemia (4 out of 81 = 4.9%). Sickle cell disease patients with a hemoglobin level above 7.5 g/dL do not need PRBCs transfusion prior to adenotonsillectomy. This approach did not increase the risk of postoperative surgical or SCD-related complications.


Clonal Diversity and Epidemiological Characteristics of ST239-MRSA Strains.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen associated with severe morbidity and mortality and poses a significant threat to public health worldwide. The genetic diversity based on sequence types of MRSA strains was illustrated in previous studies; meanwhile, the diversity along with the predominant sequence type, especially in Egypt, remains unknown. The purpose of the current study was to determine the diversity of the predominant MRSA clone ST239-MRSA (n = 50) isolated from different hosts and clinical samples and to illustrate the correlation between the resistance patterns, toxin genes, and the genetic background in Port-said and El-Sharkia Governorates, Egypt. The ST239-MRSA clone was analyzed by phenotypic antibiotyping and various genotypic assays comprising SCCmec, agr, spa, coa, and coa-RFLP in addition to toxin gene profiles. Most of the analyzed strains (40/50, 80%) were multidrug resistant (MDR), belonged to SCCmec-III, agr-I, and coa genotype I, and harbored sea and pvl genes. A negative correlation between the toxin gene profiles and antimicrobial resistance was recorded. Meanwhile, the correlation between the toxin gene profiles and the genetic background was not observed in this study. Although ST239-MRSA strains belonged to a single sequence type, they exhibited a high degree of phenotypic and genotypic diversity, indicating weak clonality and adaptability. With such diversity, it is assumed that these strains may have undergone different evolutionary processes during transmission events among and/or within a single host or tissue niche.


A test of the goodness of fit of the generic metacognitive model of psychopathology symptoms.
Common mental disorders such as depression and anxiety frequently co-occur and may share etiological mechanisms. The metacognitive model is based on the principle that there are common pathological mechanisms across disorders that account for comorbidity and therefore can be conceptualized in one generic model. A central prediction of the model is that particular metacognitive beliefs concerning the value of worry, and the uncontrollability and danger of cognition are positively correlated with psychopathology symptoms. In the present study, we set out to test the overall fit of this model by assessing generic metacognitive beliefs and judgements of attention control capacity as predictors of common and frequently co-occurring emotional distress symptoms. In a cross-sectional design, 645 participants gathered at convenience completed a battery of self-report questionnaires. Structural equation modelling indicated a good model fit for the generic metacognitive model, and the predictors accounted for 93% of the variance in distress consisting of depression-, generalized- and social anxiety symptoms. This finding supports the generic model and the implication that it can be used as a basis to formulate and treat multiple presenting problems.


Effects of surgical anaesthesia on the viability of nigral grafts in the rat striatum.
Only a small proportion of dopamine neurons in nigral grafts typically survive transplantation into the adult striatum. Since many anaesthetics reduce blood flow and disturb a variety of brain metabolites, surgical anaesthesia may be one of the factors that compromise graft survival. Conversely, the lowered core body temperature induced by some anaesthetics might promote the survival of grafted cells by slowing their metabolism. In an initial screen, the widely-used surgical anaesthetic, equithesin, was found to reduce core temperature, mean arterial blood pressure, and to increase the partial pressure of oxygen in arterial blood without producing any significant alteration in arterial pH or the partial pressure of carbon dioxide. In the main experiment, rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal bundle received dopamine-rich embryonic nigral grafts injected into the deafferented neostriatum via previously implanted guide cannulae, which allowed comparison to be made of graft survival after transplantation into awake and in re-anaesthetised animals. There were no significant differences between groups in either the functional effects of the grafts to compensate amphetamine-induced rotation, or in the survival and growth of the grafts as measured in post mortem histology. We therefore conclude that anaesthesia per se is not a major contributory factor in the relatively poor survival of dopamine neurons following transplantation into the rat striatum.


Lifestyle factors associated with incidence of rheumatoid arthritis in US adults: analysis of National Health and Nutrition Examination Survey database and meta-analysis.
To quantify rheumatoid arthritis (RA) cases attributable to selected non-genetic risk factors. National Health and Nutrition Examination Survey (NHANES) and meta-analysis. US adults. The prevalence of exposure was obtained from NHANES. Weighted analysis was performed to account for the complex sampling design in NHANES. PubMed and Web of Science up to 31 March 2019 were searched to identify epidemiological studies reported the association between non-genetic risk factors and RA in US adults. Relative risk (RR) value and the corresponding CI were pooled by meta-analysis to evaluate the associations between modifiable risk factors and RA. Population attributable fraction (PAF) was calculated based on the prevalence and RR data. The weighted percentages of former smokers, current smokers and overweight or obese people were 24.84%, 23.93% and 63.97%, and the average alcohol consumption was 51.34 g/week. In the meta-analysis, we found that former smokers (RR 1.22, 95% CI 1.10 to 1.36) and current smokers (RR 1.47, 95% CI 1.29 to 1.68) had higher risks of RA. Overweight and obese individuals had 1.27-fold (95% CI 1.09 to 1.48) increased risk of RA. Each per 50 g/week increment of alcohol consumption was associated with 8% (95% CI 0% to 16%) reduction in the risk of RA. Therefore, PAF value of smoking was 14.00% (95% CI 8.13% to 23.33%). Excess body mass index (BMI) was found to account for 14.73% (95% CI 5.45% to 23.50%) of RA incidence. The fraction of RA risk attributed by low alcohol intake was 8.21% (95% CI 0.31% to 16.39%). Collectively, we found that 32.69% (95% CI 13.41% to 50.96%) of RA cases were attributable to smoking, overweight or obesity and low alcohol drinking. Nearly 33% of RA incidence was attributed to smoking, excess BMI and low alcohol drinking in USA. Our findings could provide a basis for developing guidelines of RA prevention and control in USA.


A perspective on the economic valorization of gene manipulated biotechnology: Past and future.
Three distinct fields of gene manipulated biotechnology have so far been economically exploited: medical biotechnology, plant biotechnology and industrial biotechnology. This article analyzes the economic evolution and its drivers in the three fields over the past decades, highlighting strong divergences. Product and market characteristics, affecting firms' financing options, are shown to be important enablers or inhibitors. Subsequently, the lack of commercialization in a fourth type of gene manipulated biotechnology, namely environmental biotechnology, is explained by the existence of strong barriers. Given the latter's great promises for environmental sustainability, we argue for a need to push the commercial valorization of environmental biotechnology. Our research has strong implications for (technology) management research in biotechnology, pointing to a need to control for and/or distinguish between different biotechnology fields.


The antihypertensive hydralazine is an efficient scavenger of acrolein.
Recent work indicates the highly toxic alpha,beta-unsaturated aldehyde acrolein is formed during the peroxidation of polyunsaturated lipids, raising the possibility that it functions as a 'toxicological second messenger' during oxidative cell injury. Acrolein reacts rapidly with proteins, forming adducts that retain carbonyl groups. Damage by this route may thus contribute to the burden of carbonylated proteins in tissues. This work evaluated several amine compounds with known aldehyde-scavenging properties for their ability to attenuate protein carbonylation by acrolein. The compounds tested were: (i) the glycoxidation inhibitors, aminoguanidine and carnosine; (ii) the antihypertensive, hydralazine; and (iii) the classic carbonyl reagent, methoxyamine. Each compound attenuated carbonylation of a model protein, bovine serum albumin, during reactions with acrolein at neutral pH and 37 degrees C. However, the most efficient agent was hydralazine, which strongly suppressed carbonylation under these conditions. Study of the rate of reaction between acrolein and the various amines in a protein-free buffered system buttressed these findings, since hydralazine reacted with acrolein at rates 2-3 times faster than its reaction with the other scavengers. Hydralazine also protected isolated mouse hepatocytes against cell killing by allyl alcohol, which undergoes in situ alcohol dehydrogenase-catalysed conversion to acrolein.


Comprehensive clinical care for men who have sex with men: an integrated approach.
Men who have sex with men (MSM) have unique health-care needs, not only because of biological factors such as an increased susceptibility to infection with HIV and sexually transmitted infections associated with their sexual behaviour, but also because of internalisation of societal stigma related to homosexuality and gender non-conformity, resulting in depression, anxiety, substance use, and other adverse outcomes. Successful responses to the global HIV/AIDS epidemic will require the development of culturally sensitive clinical care programmes for MSM that address these health disparities and root causes of maladaptive behaviour (eg, societal homophobia). Health-care providers need to become familiar with local outreach agencies, hotlines, and media that can connect MSM with positive role models and social opportunities. Research is needed to understand how many MSM lead resilient and productive lives in the face of discrimination to develop assets-based interventions that build on community support. Optimum clinical care for sexual and gender minorities is a fundamental human right. MSM deserve to be treated with respect, and health-care providers need to interact with them in ways that promote disclosure of actionable health information.


Suspected bacterial endophthalmitis following sustained-release dexamethasone intravitreal implant: a case report.
A 58-year-old man admitted to our opthalmology department with the complaint of branch retinal vein occlusion. He was treated with intravitreal Ozurdex in the right eye. Two days after the injection, the patient presented with ocular pain and the visual acuity was hand movement. A diagnosis of endophthalmitis was made. We performed emergent pars plana vitrectomy (PPV) and the implant was removed from the vitreous cavity using a retinal forceps. A combination of vancomycin 1.0 mg and amikacin 0.4 mg was injected intravitreally. However, because of the blurring in the vitreus one week after the procedure, phacoemulsification and a repeat PPV was performed. Five days after the last procedure the signs and symptoms of endophthalmitis were resolved. Our case demonstrated that endophthalmitis could develop after intravitreal implantation of Ozurdex. Surgical removal of the implant and immediate vitrectomy seems to be a useful treatment option in these cases.


Intranervous immunoglobulin deposits: an underestimated mechanism of neuropathy.
There are several pathogenic mechanisms of peripheral nerve involvement in patients with monoclonal dysglobulinemia. Intranervous proliferation of malignant cells, immunoglobulin, or amyloid deposits in the endoneurial space can only be determined by examination of nerve biopsy specimens. We present clinical, electrophysiological, and histological data from seven patients whose polyneuropathy was induced by immunoglobulin deposits in the endoneurial space. As these lesions cannot be demonstrated on clinical and electrophysiological grounds, the indication for nerve biopsy derives from careful analysis of each patient presenting with a polyneuropathy and a monoclonal dysglobulinemia. To visualize and clearly characterize these deposits, electron microscopic examination is indispensable. Immunocytochemical methods using both light and electron microscopy for ultrastructural analysis are of great value. Demonstration of endoneurial immunoglobulin deposits may have major therapeutic consequences. Indeed, identification of these deposits prompted the use of aggressive treatment, which was quite effective in five of our seven patients.


Non-toxic, non-biocide-release antifouling coatings based on molecular structure design for marine applications.
Marine biofouling generally refers to the undesirable accumulation of biological organisms on surfaces in contact with seawater. This natural phenomenon represents a major economic concern for marine industries, e.g. for ships and vessels, oil and wind-turbine sea-platforms, pipelines, water valves and filters, as it limits the performance of devices, materials and underwater structures and increases the costs related to transport delays, hull maintenance and repair, cleaning and desalination units, corrosion and structure break-down. In the last few decades, many efforts have been spent into developing efficient antifouling (AF) surfaces (coatings) combining advances in materials science and recent knowledge of marine chemistry and biology. However, the extensive use of toxic and harmful compounds in the formulations raised increasing health and environmental concerns leading to stricter regulations which pushed marine industries to search for new AF strategies. This review presents the recent research progress made in green strategies for AF coatings using non-toxic, non-biocide-release based principles for marine applications. The two main approaches, detachment of biofoulants or preventing biofoulants attachment, are reviewed in detail and new promising routes based on amphiphilic, (super)hydrophilic, and topographic (structured) surfaces are highlighted. The chemical and physical aspects of the AF mechanisms behind the AF strategies reviewed are emphasized, with special attention to the early stages of biofoulant adhesion, keeping the focus on the materials' molecular structure and properties which allow obtaining the final desired antifouling behaviour.


Genetic influence on early age-related maculopathy: a twin study.
Age-related macular degeneration (AMD) is the most common cause of blindness in industrialized countries. There has been considerable interest in the genetics of early age-related maculopathy (ARM) and AMD, because they have phenotypes similar to inherited diseases where mutations have been identified, but the heritability of ARM and AMD is unknown. A classical twin study was performed to compare the concordance in monozygotic (MZ) and dizygotic (DZ) twins in an unselected sample of female volunteer twins. Five hundred six twin pairs, 226 MZ and 280 DZ, with a mean age of 62 years, were examined. ARM was graded from stereoscopic macular photographs of 501 of the twin pairs (99%) according to the International ARM Epidemiologic Study Group grading system. The casewise concordance was calculated for twin pairs from 2 x 2 contingency tables of affected/unaffected twins, and these tables were used in maximum likelihood genetic modeling to estimate the heritabilities of phenotypes graded. Prevalence of ARM; concordance in MZ and DZ twins of the phenotypes of ARM, soft drusen >63 microm and > or =125 microm diameter, pigmentary changes and hard drusen (<20 and > or =20 in number); heritability of ARM and subphenotypes. The overall prevalence of ARM was 14.6% (95% confidence interval [CI], 12.4%-16.8%). The concordance for ARM in MZ twins was 0.37 compared with 0.19 in DZ twins, suggesting a role for genes. Modeling confirmed a genetic effect for phenotypes of ARM, soft drusen, pigmentary changes, and > or =20 hard drusen, although there was little genetic effect for scattered (<20) hard drusen. The heritability of ARM was estimated as 45% (95% CI, 35%-53%). The most heritable phenotypes were soft drusen > or =125 microm (57%) and > or =20 hard drusen (81%), with the latter being dominantly inherited. This study confirms a significant genetic influence in ARM and suggests that future genetic studies should examine phenotypes of large (> or =125 microm) soft drusen and > or =20 hard drusen, because these seem to be the most heritable components.


CREB is a critical regulator of normal hematopoiesis and leukemogenesis.
The cAMP-responsive element binding protein (CREB) is a 43-kDa nuclear transcription factor that regulates cell growth, memory, and glucose homeostasis. We showed previously that CREB is amplified in myeloid leukemia blasts and expressed at higher levels in leukemia stem cells from patients with myeloid leukemia. CREB transgenic mice develop myeloproliferative disease after 1 year, but not leukemia, suggesting that CREB contributes to but is not sufficient for leukemogenesis. Here, we show that CREB is most highly expressed in lineage negative hematopoietic stem cells (HSCs). To understand the role of CREB in hematopoietic progenitors and leukemia cells, we examined the effects of RNA interference (RNAi) to knock down CREB expression in vitro and in vivo. Transduction of primary HSCs or myeloid leukemia cells with lentiviral CREB shRNAs resulted in decreased proliferation of stem cells, cell- cycle abnormalities, and inhibition of CREB transcription. Mice that received transplants of bone marrow transduced with CREB shRNA had decreased committed progenitors compared with control mice. Mice injected with Ba/F3 cells expressing either Bcr-Abl wild-type or T315I mutation with CREB shRNA had delayed leukemic infiltration by bioluminescence imaging and prolonged median survival. Our results suggest that CREB is critical for normal myelopoiesis and leukemia cell proliferation.


Short-term effects of marathon running: no evidence of cardiac dysfunction.
The purpose of this study was to analyze the short-term effects of a marathon race (Madrid Marathon) on both markers of cardiac damage and echocardiographic parameters in a group of 22 runners (17 male and 5 female; 34 +/- 5 yr; VO2max: 55.7 +/- 9.1 mL x kg(-1) x min(-1) with a wide range of fitness levels. Venous blood samples were collected from each subject 48 h before the race, at race finish, and 6, 24, and 48 h postexercise for the determination of myoglobin, total creatine kinase catalytic activity (total CK), mass concentration of creatine kinase isoenzyme MB (CK-MB mass), and cardiac isoforms of troponin T and I (TnT-c and TnI-c, respectively). In addition, echocardiographic parameters (M-mode two-dimensional and Doppler analysis) indicative of both left ventricular (LV) systolic and diastolic function were obtained three times from each runner: 2-5 d before the race, at race finish, and 24-36 h after exercise. Except in one subject, levels of TnT-c and TnI-c were within normal limits (<0.1 ng x mL(-1)) in all the samples collected before or after the race. Overall LV systolic function was not altered by marathon running. Finally, LV diastolic function was transiently altered after the race since the ratio between peak early and late transmitral filling velocities (E/A) was significantly reduced at race finish (P < 0.01) and returned to resting levels after 24-36 h. Our findings suggest that marathon running does not adversely affect the hearts of healthy individuals independently from their training status.


Catastrophic neurological complications of emergent endotracheal intubation: report of 2 cases.
Although exceedingly rare, catastrophic neurological decline may result from endotracheal intubation of patients with preexisting cervical spine disease. The authors report on 2 cases of quadriplegia resulting from emergent endotracheal intubation in the intensive care unit. A 68-year-old man with ankylosing spondylitis became quadriplegic after emergent intubation. A new C6-7 fracturedislocation was identified, and the patient underwent emergent open reduction and C4-T2 posterior fixation and fusion. The patient remained quadriplegic and ultimately died of pneumonia 1 year later. This is the first report with radiographic documentation of a cervical fracture-dislocation resulting from intubation in a patient with ankylosing spondylitis. A 73-year-old man underwent posterior C6-T1 decompression and fixation for a C6-7 fracture. On postoperative Day 12, emergent intubation for respiratory distress resulted in C6-level quadriplegia. Imaging revealed acute spondyloptosis at C6-7, and the patient underwent emergent open reduction with revision and extension of posterior fusion from C-3 to T-2. He remained quadriplegic and ventilator dependent. Five days after the second operation, care was withdrawn. This is the first report of intubation as a cause of significant neurological decline related to disruption of a recently fixated cervical fracture. Risk factors are identified and pertinent literature is reviewed for cases of catastrophic neurological complications after emergent endotracheal intubation. Strategies for obtaining airway control in patients with cervical spine pathology are also identified. Awareness of the potential dangers of airway management in patients with cervical spine pathology is critical for all involved subspecialty team members.


H19 and MEST gene expression and histone modification in blastocysts cultured from vitrified and fresh two-cell mouse embryos.
Vitrification of embryos is a routine procedure in many IVF laboratories. The effect of vitrification on gene expression and some modifications of H3 histone in H19 and MEST imprinted genes in blastocysts produced in vitro from non-vitrified and vitrified two-cell embryos was investigated. The expression level of the chosen imprinted genes increased significantly (P < 0.05) in experimental groups compared with in-vivo blastocysts (control group). H3K9me2 decreased, whereas H3K9ac increased in the experimental group compared with the control group. The increases in the expression levels of the imprinted genes, and the attendant changes in histone and chromatin status associated with in-vitro culture of embryos from the two-cell stage, are unaffected by prior vitrification and warming. In the present study, it was shown that such changes are solely caused by the effect of in-vitro culture, irrespective of vitrification. Although these genes are sensitive to environmental changes, vitrification seems to have no additional effect on these genes and on the histone marks, and can threfore be considered to be a process with minimum damage for embryo cryopreservation in assisted reproductive technology applications.


Mechanisms of lymphatic regeneration after tissue transfer.
Lymphedema is the chronic swelling of an extremity that occurs commonly after lymph node resection for cancer treatment. Recent studies have demonstrated that transfer of healthy tissues can be used as a means of bypassing damaged lymphatics and ameliorating lymphedema. The purpose of these studies was to investigate the mechanisms that regulate lymphatic regeneration after tissue transfer. Nude mice (recipients) underwent 2-mm tail skin excisions that were either left open or repaired with full-thickness skin grafts harvested from donor transgenic mice that expressed green fluorescent protein in all tissues or from LYVE-1 knockout mice. Lymphatic regeneration, expression of VEGF-C, macrophage infiltration, and potential for skin grafting to bypass damaged lymphatics were assessed. Skin grafts healed rapidly and restored lymphatic flow. Lymphatic regeneration occurred beginning at the peripheral edges of the graft, primarily from ingrowth of new lymphatic vessels originating from the recipient mouse. In addition, donor lymphatic vessels appeared to spontaneously re-anastomose with recipient vessels. Patterns of VEGF-C expression and macrophage infiltration were temporally and spatially associated with lymphatic regeneration. When compared to mice treated with excision only, there was a 4-fold decrease in tail volumes, 2.5-fold increase in lymphatic transport by lymphoscintigraphy, 40% decrease in dermal thickness, and 54% decrease in scar index in skin-grafted animals, indicating that tissue transfer could bypass damaged lymphatics and promote rapid lymphatic regeneration. Our studies suggest that lymphatic regeneration after tissue transfer occurs by ingrowth of lymphatic vessels and spontaneous re-connection of existing lymphatics. This process is temporally and spatially associated with VEGF-C expression and macrophage infiltration. Finally, tissue transfer can be used to bypass damaged lymphatics and promote rapid lymphatic regeneration.


ER and PR immunohistochemistry and HER2 FISH versus oncotype DX: implications for breast cancer treatment.
Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor (HER2) concordance between immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), and Oncotype DX, a commercially available RT-PCR-based assay which recently began reporting biomarker results was assessed. ER concordance was 98.9% (262/265), Pearson correlation coefficient (r) = 0.42, and Spearman's rank correlation (ρ) = 0.25. Positive percent agreement for ER was 98.9% (262/265). One patient with discordant ER results was not offered hormone therapy based on the preferential use of Oncotype DX. PR was concordant in 91.3% (242/265), r = 0.80, ρ = 0.75, and Cohen's kappa (κ) = 0.63. Positive percent agreement for PR was 90.5% (218/241) and negative percent agreement was 100% (24/24). HER2 concordance was 99.2% (245/247), r = 0.35, ρ = 0.28, and κ = 0.12. Positive percent agreement for HER2 was 0% (0/2) and negative percent agreement was 100% (245/245). Of the three FISH HER2-amplified cases, two were negative and one was equivocal, and all FISH HER2-equivocal cases (n = 3) were negative by Oncotype DX. Patients that were FISH HER2-amplified, Oncotype DX HER2-negative did not receive trastuzumab. Although our results demonstrated high concordance between IHC and Oncotype DX for ER and PR, our data showed poor positive percent agreement for HER2. Compared to FISH, Oncotype DX does not identify HER2-positive breast carcinomas. The preferential use of Oncotype DX biomarker results over IHC and FISH is discouraged.


An expanded RT-PCR melting temperature coding assay to rapidly identify all known SARS-CoV-2 variants and sub-variants of concern.
The continued emergence of vaccine-resistant SARS-CoV-2 variants of concern (VOC) requires specific identification of each VOC as it arises. Here, we report an expanded version of our previously described sloppy molecular beacon (SMB) melting temperature (Tm) signature-based assay for VOCs, now modified to include detection of Delta (B.1.617.2) and Omicron (B.1.1.529) sub-variants. The SMB-VOC assay targets the signature codons 501, 484 and 452 in the SARS-CoV-2 spike protein which we show can specifically detect and differentiate all known VOCs including the Omicron subvariants (BA.1, BA.2, BA.2.12.1, BA.4/BA.5). The limit of detection (LOD) of the assay was 20, 22 and 36 genomic equivalents (GE) per reaction with the Delta, Omicron BA.1 and BA.2 respectively. Clinical validation of the 3-codon assay in the LC480 instrument showed the assay detected 94% (81/86) of the specimens as WT or VOCs and 6% (5/86) of the tests producing indeterminate results compared to sequencing. Sanger sequencing also failed for four samples. None of the specimens were incorrectly identified as WT or as a different VOC by our assay. Thus, excluding specimens with indeterminant results, the assay was 100% sensitive and 100% specific compared to Sanger sequencing for variant identification. This new assay concept can be easily expanded to add newer variants and can serve as a robust diagnostic tool for selecting appropriate monoclonal antibody therapy and rapid VOC surveillance.


Effects of GH/IGF on the Aging Mitochondria.
The mitochondria are key organelles regulating vital processes in the eukaryote cell. A decline in mitochondrial function is one of the hallmarks of aging. Growth hormone (GH) and the insulin-like growth factor-1 (IGF-1) are somatotropic hormones that regulate cellular homeostasis and play significant roles in cell differentiation, function, and survival. In mammals, these hormones peak during puberty and decline gradually during adulthood and aging. Here, we review the evidence that GH and IGF-1 regulate mitochondrial mass and function and contribute to specific processes of cellular aging. Specifically, we discuss the contribution of GH and IGF-1 to mitochondrial biogenesis, respiration and ATP production, oxidative stress, senescence, and apoptosis. Particular emphasis was placed on how these pathways intersect during aging.


Inflammatory priming predisposes mice to age-related retinal degeneration.
Disruption of cellular processes affected by multiple genes and accumulation of numerous insults throughout life dictate the progression of age-related disorders, but their complex etiology is poorly understood. Postmitotic neurons, such as photoreceptor cells in the retina and epithelial cells in the adjacent retinal pigmented epithelium, are especially susceptible to cellular senescence, which contributes to age-related retinal degeneration (ARD). The multigenic and complex etiology of ARD in humans is reflected by the relative paucity of effective compounds for its early prevention and treatment. To understand the genetic differences that drive ARD pathogenesis, we studied A/J mice, which develop ARD more pronounced than that in other inbred mouse models. Although our investigation of consomic strains failed to identify a chromosome associated with the observed retinal deterioration, pathway analysis of RNA-Seq data from young mice prior to retinal pathological changes revealed that increased vulnerability to ARD in A/J mice was due to initially high levels of inflammatory factors and low levels of homeostatic neuroprotective factors. The genetic signatures of an uncompensated preinflammatory state and ARD progression identified here aid in understanding the susceptible genetic loci that underlie pathogenic mechanisms of age-associated disorders, including several human blinding diseases.


Pushing and pulling in prokaryotic DNA segregation.
In prokaryotes, DNA can be segregated by three different types of cytoskeletal filaments. The best-understood type of partitioning (par) locus encodes an actin homolog called ParM, which forms dynamically unstable filaments that push plasmids apart in a process reminiscent of mitosis. However, the most common type of par locus, which is present on many plasmids and most bacterial chromosomes, encodes a P loop ATPase (ParA) that distributes plasmids equidistant from one another on the bacterial nucleoid. A third type of par locus encodes a tubulin homolog (TubZ) that forms cytoskeletal filaments that move rapidly with treadmill dynamics.


The Role of 17β-Estradiol and Estrogen Receptors in Regulation of Ca2+ Channels and Mitochondrial Function in Cardiomyocytes.
Numerous epidemiological, clinical, and animal studies showed that cardiac function and manifestation of cardiovascular diseases (CVDs) are different between males and females. The underlying reasons for these sex differences are definitely multifactorial, but major evidence points to a causal role of the sex steroid hormone 17β-estradiol (E2) and its receptors (ER) in the physiology and pathophysiology of the heart. Interestingly, it has been shown that cardiac calcium (Ca2+) ion channels and mitochondrial function are regulated in a sex-specific manner. Accurate mitochondrial function and Ca2+ signaling are of utmost importance for adequate heart function and crucial to maintaining the cardiovascular health. Due to the highly sensitive nature of these processes in the heart, this review article highlights the current knowledge regarding sex dimorphisms in the heart implicating the importance of E2 and ERs in the regulation of cardiac mitochondrial function and Ca2+ ion channels, thus the contractility. In particular, we provide an overview of in-vitro and in-vivo studies using either E2 deficiency; ER deficiency or selective ER activation, which suggest that E2 and ERs are strongly involved in these processes. In this context, this review also discusses the divergent E2-responses resulting from the activation of different ER subtypes in these processes. Detailed understanding of the E2 and ER-mediated molecular and cellular mechanisms in the heart under physiological and pathological conditions may help to design more specifically targeted drugs for the management of CVDs in men and women.


[Diffuse deep cervical infection. Report of three cases].
Deep cervical infections still appear despite antibiotic use. Necrotizing fasciitis of the head and neck is a rare but serious infection caused by mixed, predominantly anaerobic bacterial flora. The natural course of the disease is spreading necrosis of the soft tissues of the neck and septicemia. Three recent cases of cervical necrotizing soft-tissue infection are presented. The importance of early and aggressive surgical treatment complemented by parenteral antibiotics with an aerobic-anaerobic spectrum is emphasized.


Local plasticity of dendritic excitability can be autonomous of synaptic plasticity and regulated by activity-based phosphorylation of Kv4.2.
While plasticity is typically associated with persistent modifications of synaptic strengths, recent studies indicated that modulations of dendritic excitability may form the other part of the engram and dynamically affect computational processing and output of neuronal circuits. However it remains unknown whether modulation of dendritic excitability is controlled by synaptic changes or whether it can be distinct from them. Here we report the first observation of the induction of a persistent plastic decrease in dendritic excitability decoupled from synaptic stimulation, which is localized and purely activity-based. In rats this local plasticity decrease is conferred by CamKII mediated phosphorylation of A-type potassium channels upon interaction of a back propagating action potential (bAP) with dendritic depolarization.


How analgesia-sedation alters monocyte mCD14 and HLA-DR levels in patients undergoing cerebral angiography?
This study aims to evaluate how analgesia-sedation causes alterations of HLA-DR and cytokines (IL-10 and IL-6) in patients undergoing cerebral angiography for several cerebral vascular diseases. This study includes 41 males who underwent cerebral angiography. The study sample was divided into two: Group I had 7 patients who did not receive and group II had 34 patients who received analgesia-sedation before cerebral angiography. A venous blood sample was collected before and after cerebral angiography. Analgesia-sedation caused significant increase CD4+ and CD19 T lymphocytes (p < 0.001) but group I showed significant increase in CD40, CD154, and MHC-II levels (p < 0.001) after cerebral angiography. We suggest that the effects of fentanyl and midazolam on the immune response are the reflection of the effects by the monocyte, mHLA-DR expression. In the future, depending on the immunological status of the patients, different anesthetic applications including the new anesthetic agents that will be able to decrease immune system suppression will be required.


Activation of estrogen receptor-alpha by the heavy metal cadmium.
Previous studies from this laboratory have shown that the heavy metal cadmium (Cd) mimics the effects of estradiol in estrogen-responsive breast cancer cell lines. To understand the mechanism by which cadmium activates estrogen receptor-alpha (ER-alpha), the ability of cadmium to bind to and activate wild-type and various mutants of ER-alpha was examined. When tested in transient cotransfection assays in COS-1 cells, cadmium concentrations as low as 10(-11) M activated ER-alpha. Scatchard analysis employing either purified human recombinant ER-alpha or extracts from ER-containing MCF-7 cells demonstrated that l09Cd binds to the ER with an equilibrium dissociation constant of approximately 4 to 5 x 10(-10) M. Cadmium also blocks the binding of estradiol to ER-alpha in a noncompetitive manner (K(i) = 2.96 x 10(-10) M), suggesting that the heavy metal interacts with the hormone-binding domain of the receptor. To study the role of the hormone-binding domain in cadmium activation, COS-1 cells were transiently cotransfected with GAL-ER, a chimeric receptor containing the DNA-binding domain of the transcription factor GAL4 and the hormone-binding domain of ER-alpha, and a GAL4-responsive reporter gene. Treatment of the transfected cells with either 10(-6) M cadmium or 10(-9) M estradiol resulted in a 4-fold increase in reporter gene activity. The effect of cadmium on the chimeric receptor was blocked by the antiestrogen, ICI-164,384, suggesting that cadmium activates ER-alpha through an interaction with the hormone-binding domain of the receptor. Transfection and binding assays with ER-alpha mutants identified C381, C447, E523, H524, and D538 as possible interaction sites of cadmium with the hormone-binding domain of ER-alpha.


Neural plasticity: Part 3. Responses to lesions in the peripheral nervous system.
This paper describes some of the remarkable recuperative responses that neurons of the peripheral nervous system have to injury of their axons. The initial response is a sealing of the cut ends of the axons, followed by orderly degenerative and phagocytic events in the segments lying distal and proximal to the lesion. The chromatolytic changes occurring in the cell body are adaptive responses involving the morphological, physiological, and biochemical properties of the neuron. The cell loses dendrites and synapses, its electrophysiological properties are altered, and its biochemistry is altered to prepare the neuron for synthesizing proteins needed for formation of growth cones and regeneration of the axon. During this period, the denervated muscle undergoes dramatic atrophic changes that not only modify the electrical nd chemical properties of its membrane but also reduce the diameter and weight of its fibers. This denervation atrophy of muscle is reversible if reinnervation occurs before muscle material has been replaced by connective tissue. Responses to injury of sensory axons depend on the site of the lesion. Cutting a dorsal root causes Wallerian degeneration of the primary afferents' central processes. Collateral sprouts from neighboring uncut root fibers are thought to take over the vacant synapses on the second-order neurons in the dorsal horn. Cutting a sensory axon peripherally eliminates the receptive field of the primary afferent initially, but in time, collaterals from neighboring skin afferents provide the second-order neurons with a new receptive field. Synaptic reclamation and receptive field reorganization are but two dramatic examples of neuronal plasticity that lead to "rewired" neural circuitry.


Distribution of basic fibroblast growth factor in the 18-day rat fetus: localization in the basement membranes of diverse tissues.
Immunohistochemical methods were used to study the distribution of basic FGF in the 18-d rat fetus. The results reveal a pattern of widespread yet specific staining that is consistent with the wide distribution of basic FGF. Immunoreactive basic FGF is associated with mesenchymal structures, mesoderm- and neuroectoderm-derived cells, and their extracellular matrices. As an example, skeletal and smooth muscle cells are strongly positive. The basement membrane underlying the epithelia always contain basic FGF. In some tissues (i.e., cartilage and bone) the intensity of immunostaining is dependent on the stage of cell differentiation. Although the staining of tissues is primarily associated with the extracellular matrix, there is significant intracellular staining in various cell types. This is particularly evident in the endocrine cells of the adrenal cortex, testis, and ovary. The histochemical findings reported here support the notion that basic FGF has the characteristics required to mediate many of the effects of the mesenchyme on cell growth and differentiation. The significance of these findings in understanding the role of basic FGF in regulating cell proliferation and differentiation is discussed.


Laterality, hand control and scholastic performance: a British birth cohort study.
To use simple measures of laterality and hand control that can identify a greater risk of poorer scholastic ability, potentially signalling suboptimal hemispheric lateralisation. Analysis of material from a birth cohort study. Members of the National Child Development Study, a British birth cohort study following people born in 1958. 10 612 children who undertook tests at age 11 years. Teacher-administered tests of non-verbal general ability, verbal general ability, reading comprehension and mathematics. Linear regression produced associations (and 95% CIs) with tests of verbal general ability, non-verbal general ability, reading comprehension and mathematics scores for the lowest third (compared with highest) of a left-hand control test involving picking up matches of -1.21 (-1.73 to -0.68; p<0.001), -0.72 (-1.14 to -0.29; p=0.001), -0.70 (-1.06 to -0.35; p<0.001) and -1.32 (-1.90 to -0.73; p<0.001). Among those in the lowest third of the right-hand control test score, mixed-handedness compared with right-handedness was associated with poorer scholastic performance, with regression coefficients (and 95% CIs; p values) of 1.90 (-3.01 to -0.80; p=0.001), -1.25 (-2.15 to -0.35; p=0.007), -1.28 (2.04 to -0.53; p=0.001) and -1.33 (-2.53 to -0.13; p=0.030). The estimates are for a point change in the scholastic test scores, after adjustment for sex, left-hand motor function and social class. Statistically significant associations with mixed-handedness were only observed for the lowest third of right-hand motor function. Measures involving poorer left-hand motor function may represent useful markers of reduced cognitive function possibly reflecting suboptimal hemispheric lateralisation. Crude measures of laterality such as reported non-right-handedness may be more useful for research when combined with measures of motor function.


Isolation and characterization of a staphylococcal lipase.
A number of coagulase-negative staphylococci isolated from human skin were found to produce lipase. Lipolytic activity appeared in the growth medium during the stationary phase of growth but did not appear as a result of autolysis of the cells. Maximal lipase synthesis was obtained when the medium was adjusted to pH 7.5 before inoculation. The purified enzyme hydrolyzed tributyrin and tridecanoin most actively, and a relatively high rate of hydrolysis of triolein was also noted. The optimal activity of the purified lipase was at pH 7.5. The characteristics of the concentrated crude enzyme and purified lipase were compared.


[Severe thromboembolic complications following lumbar spinal phlebography].
From a series of 400 cases of lumbar spine phlebography by selective catheterisation, 4 serious thrombo-embolic episodes were observed, including 2 cases of severe pulmonary embolism which responded favourably to medical treatment. In 3 cases, risk factors were found (prolonged immobilisation in bed past history of phlebitis). One case developed severe phlebitis of the lower limbs despite prophylactic treatment with low dose calcium heparin. In the other cases, no anticoagulant therapy was prescribed before or during the examination.


Prognostic and therapeutic relevance of molecular subtypes in high-grade serous ovarian cancer.
Molecular classification of high-grade serous ovarian cancer (HGSOC) using transcriptional profiling has proven to be complex and difficult to validate across studies. We determined gene expression profiles of 174 well-annotated HGSOCs and demonstrate prognostic significance of the prespecified TCGA Network gene signatures. Furthermore, we confirm the presence of four HGSOC transcriptional subtypes using a de novo classification. Survival differed statistically significantly between de novo subtypes (log rank, P = .006) and was the best for the immunoreactive-like subtype, but statistically significantly worse for the proliferative- or mesenchymal-like subtypes (adjusted hazard ratio = 1.89, 95% confidence interval = 1.18 to 3.02, P = .008, and adjusted hazard ratio = 2.45, 95% confidence interval = 1.43 to 4.18, P = .001, respectively). More prognostic information was provided by the de novo than the TCGA classification (Likelihood Ratio tests, P = .003 and P = .04, respectively). All statistical tests were two-sided. These findings were replicated in an external data set of 185 HGSOCs and confirm the presence of four prognostically relevant molecular subtypes that have the potential to guide therapy decisions.


Stagnant Physical Therapy Referral Rates Alongside Rising Opioid Prescription Rates in Patients With Low Back Pain in the United States 1997-2010.
A cross-sectional observational study utilizing the National Ambulatory and National Hospital Ambulatory Medical Care Surveys between 1997 and 2010. The aim of this study was to characterize national physical therapy (PT) referral trends during primary care provider (PCP) visits in the United States. Despite guidelines recommending PT for the initial management of low back pain (LBP), national PT referral rates remain low. Race, ethnicity, age, payer type, and PT referral rates were collected for patients aged 16 to 90 years who were visiting their PCPs. Associations among demographic variables and PT referral were determined using logistic regression. Between 1997 and 2010, we estimated 170 million visits for LBP leading to 17.1 million PT referrals. Average proportion of PCP visits associated with PT referrals remained stable at about 10.1% [odds ratio (OR) 1.00, 95% confidence interval (95% CI) 0.96-1.04)], despite our prior finding of increasing number of visits associated with opioid prescriptions in the same timeframe.Lower PT referral rates were observed among visits by patients who were insured by Medicaid (OR 0.48, 95% CI 0.33-0.69) and Medicare (OR 0.50, 95% CI 0.35-0.72). Furthermore, visits not associated with PT referrals were more likely to be associated with opioid prescriptions (OR 1.69, 95% CI 1.22-2.35). Although therapies delivered by PTs are promoted as a first-line treatment for LBP, PT referral rates remain low. There also exist disparately lower referral rates in populations with more restrictive health plans and simultaneous opioid prescription. Our findings provide a broad overview to PT prescription trend and isolate concerning associations requiring further explorations. 3.


Low oxygen tension and relative defined culture medium with 3, 4-dihydroxyflavone are beneficial for yak-bovine interspecies somatic cell nuclear transfer embryo.
With an aim to improve the efficiency of yak-bovine interspecies somatic cell nuclear transfer (iSCNT), this study investigated the effect of different culture systems on the development, quality and gene expression profile of yak-bovine iSCNT embryo. Reconstructed embryos were cultured in modified synthetic oviductal fluid (mSOF) or relative defined culture medium (RDCM) with 5% or 20% oxygen tension. Relative mRNA abundance of Oct-4, IFNT, IGF-2, Bax, GPX-1, SOD-1, CAT and GSS was analysed in blastocysts with qRT-PCR. The blastocyst formation rate in RDCM under 5% oxygen tension was significantly higher than that under 20% oxygen tension (P < 0.05). The total cell number of blastocyst derived from RDCM with 20% oxygen tension was lower than that of other groups, whereas the group of RDCM with 5% oxygen tension showed a beneficial effect on apoptosis index and tolerance to cryopreservation (P < 0.05). However, under the same oxygen tension, the mRNA abundance of IFNT of RDCM groups was higher than that of the mSOF groups. In addition, high oxygen tension during in vitro culture (IVC) with RDCM significantly increases the mRNA expression of oxidative stress-related genes (GPX-1, SOD-1, CAT and GSS) (P < 0.05). 3, 4-Dihydroxyflavone (DHF) during high oxygen tension was able to improve the cloned blastocyst formation rate in RDCM (P < 0.05). These results for the first time showed that low oxygen tension and RDCM could improve the developmental competence and quality and alleviate the oxidative stress for yak-bovine iSCNT embryo during IVC.


Overall C as a measure of discrimination in survival analysis: model specific population value and confidence interval estimation.
The assessment of the discrimination ability of a survival analysis model is a problem of considerable theoretical interest and important practical applications. This issue is, however, more complex than evaluating the performance of a linear or logistic regression. Several different measures have been proposed in the biostatistical literature. In this paper we investigate the properties of the overall C index introduced by Harrell as a natural extension of the ROC curve area to survival analysis. We develop the overall C index as a parameter describing the performance of a given model applied to the population under consideration and discuss the statistic used as its sample estimate. We discover a relationship between the overall C and the modified Kendall's tau and construct a confidence interval for our measure based on the asymptotic normality of its estimate. Then we investigate via simulations the length and coverage probability of this interval. Finally, we present a real life example evaluating the performance of a Framingham Heart Study model.


Alcohol and cannabis use among adolescents in Flemish secondary school in Brussels: effects of type of education.
Research regarding socio-economic differences in alcohol and drug use in adolescence yields mixed results. This study hypothesizes that (1) when using education type as a proxy of one's social status, clear differences will exist between students from different types of education, regardless of students' familial socio-economic background; (2) and that the effects of education type differ according to their cultural background. Data from the Brussels youth monitor were used, a school survey administered among 1,488 adolescents from the 3rd to 6th year of Flemish secondary education. Data were analyzed using multilevel logistic regression models. Controlling for their familial background, the results show that native students in lower educational tracks use alcohol and cannabis more often than students in upper educational tracks. Such a relationship was not found for students from another ethnic background. Results from this study indicate that research into health risks should take into account both adolescents' familial background and individual social position as different components of youngsters' socio-economic background.


Ultrafast dynamics of fluctuations in high-temperature superconductors far from equilibrium.
Despite extensive work on high-temperature superconductors, the critical behavior of an incipient condensate has so far been studied exclusively under equilibrium conditions. Here, we excite Bi(2)Sr(2)CaCu(2)O(8+δ) with a femtosecond laser pulse and monitor the subsequent nonequilibrium dynamics of the midinfrared conductivity. Our data allow us to discriminate temperature regimes where superconductivity is either coherent, fluctuating or vanishingly small. Above the transition temperature T(c), we make the striking observation that the relaxation to equilibrium exhibits power-law dynamics and scaling behavior, both for optimally and underdoped superconductors. Our findings can in part be modeled using time-dependent Ginzburg-Landau theory, and they provide strong indication of universality in systems far from equilibrium.


Aging-Induced Brain-Derived Neurotrophic Factor in Adipocyte Progenitors Contributes to Adipose Tissue Dysfunction.
Aging-related adipose tissue dysfunction contributes to the progression of chronic metabolic diseases. We investigated the role of age-dependent expression of a neurotrophin, brain-derived neurotrophic factor (BDNF) in adipose tissue. Pro-BDNF expression was elevated in epididymal white adipose tissue (eWAT) with advanced age, which was associated with the reduction in sympathetic innervation. Interestingly, BDNF expression was enriched in PDGFRα+ adipocyte progenitors isolated from eWAT, with age-dependent increase in expression. In vitro pro-BDNF treatment caused apoptosis in adipocytes differentiated from C3H10T1/2 cells, and siRNA knockdown of sortilin mitigated these effects. Tamoxifen-inducible PDGFRα+ cell-specific deletion of BDNF (BDNFPdgfra KO) reduced pro-BDNF expression in eWAT, prevented age-associated declines in sympathetic innervation and mitochondrial content in eWAT, and improved insulin sensitivity. Moreover, BDNFPdgfra KO mice showed reduced expression of aging-induced inflammation and senescence markers in eWAT. Collectively, these results identified the upregulation of pro-BDNF expression in adipocyte progenitors as a feature of visceral white adipose tissue aging and suggested that inhibition of BDNF expression in adipocyte progenitors is potentially beneficial to prevent aging-related adipose tissue dysfunction.


Synthesis and biochemical evaluation of des-vinyl secologanin aglycones with alternate stereochemistry.
Based on the X-ray structure of the enzyme strictosidine synthase, the glucose moiety of the seco-iridoid glucoside, secologanin, appears to be the key for orienting the substrate. We hypothesized that removing the glucose moiety would allow alternate stereoisomers of secologanin to be turned over. A convenient synthesis to prepare stereoisomers of des-vinyl secologanin is presented. The choice of protective group was the key to access this series of compounds. The analogs were assayed with strictosidine synthase and, interestingly, both the natural 2,4-trans diastereomer and the unnatural 2,4-cis diastereomer are turned over. The trans/cis selectivity increases with increased acetal substituent size. The results add to our understanding of how strictosidine synthase discriminates among stereoisomers.


Parathyroid hormone inhibition of phosphate transport in renal brush border vesicles from phosphate-depleted dogs.
Dietary phosphate (Pi) restriction increases renal Pi reabsorption and induces resistance to the phosphaturic action of parathyroid hormone. Na+-gradient-stimulated Pi transport in membrane vesicles isolated from the renal brush border of experimental animals has been shown to parallel changes in renal Pi reabsorption induced by dietary Pi restriction and in vivo administration of parathyroid hormone. Dietary Pi restriction has been shown to markedly inhibit the phosphaturic response to parathyroid hormone in rats and dogs. Parathyroid hormone has been reported not to decrease the Na+-gradient-stimulated transport of Pi in brush border membrane vesicles isolated from dietary Pi restricted rats unless the rats were administered an acute Pi load prior to killing, however, thyroparathyroidectomy of rats fed a low Pi diet has been reported to increase Na+-gradient-stimulated Pi transport. Using the dietary Pi restricted dog, we demonstrated no significant decrease in renal reabsorption of Pi in response to parathyroid hormone administration. However, significant decreases in Pi transport in brush border membrane vesicles isolated from the kidneys of dietary Pi restricted dogs were observed in response to in vivo parathyroid hormone administration. These data demonstrate that the resistance to the phosphaturic action of parathyroid hormone observed in vivo does not include resistance to the inhibitory effect of parathyroid hormone on Pi transport in brush border membrane vesicles. Thus, the data suggest that parathyroid hormone continues to alter Pi transport characteristics of the brush border membrane in states of Pi depletion despite the resistance to parathyroid hormone seen in vivo.


In vitro interactions of biomedical polyurethanes with macrophages and bacterial cells.
Three commercial medical-grade polyurethanes (PUs), a poly-ether-urethane (Pellethane), and two poly-carbonate-urethanes, the one aromatic (Bionate) and the other aliphatic (Chronoflex), were tested for macrophages and bacterial cells adhesion, in the presence or absence of adhesive plasma proteins. All the experiments were carried out on PUs films obtained by solvent casting. The wettability of these films was analysed by measuring static contact angles against water. The ability of the selected PUs to adsorb human fibronectin (Fn) and fibrinogen (Fbg) was checked by ELISA with biotin-labelled proteins. All PUs were able to adsorb Fn and Fbg (Fn > Fbg). Fn adsorption was in the order: Pellethane > Chronoflex > Bionate, the highest Fbg adsorption being detected onto Bionate (Bionate > Chronoflex > Pellethane). The human macrophagic line J111, and the two main bacterial strains responsible for infection in humans (Staphylococcus aureus Newman and Staphylococcus epidermidis 14852) were incubated in turn with the three PUs, uncoated or coated with plasma proteins. No macrophage or bacterial adhesion was observed onto uncoated PUs. PUs coated with plasma, Fn or Fbg promoted bacterial adhesion (S. aureus > S. epidermidis), whereas macrophage adhered more onto PUs coated with Fn or plasma. The coating with Fbg did not promote cell adhesion. Pellethane showed the highest macrophage activation (i.e. spreading), followed, in the order, by Bionate and Chronoflex.


Bridging the Gap in Global Advanced Radiation Oncology Training: Impact of a Web-Based Open-Access Interactive Three-Dimensional Contouring Atlas on Radiation Oncologist Practice in Russia.
Radiation oncologists in Russia face a number of unique professional difficulties including lack of standardized training and continuing medical education. To combat this, under the auspices of the Russian Society of Clinical Oncology (RUSSCO), our group has developed a series of ongoing in-person interactive contouring workshops that are held during the major Russian oncology conferences in Moscow, Russia. Since November 2016 during each workshop, we utilized a web-based open-access interactive three-dimensional contouring atlas as part of our didactics. We sought to determine the impact of this resource on radiation oncology practice in Russia. We distributed an IRB-approved web-based survey to 172 practicing radiation oncologists in Russia. We inquired about practice demographics, RUSSCO contouring workshop attendance, and the clinical use of open-access English language interactive contouring atlas (eContour). The survey remained open for 2 months until November 2017. Eighty radiation oncologists completed the survey with a 46.5% response rate. Mean number of years in practice was 13.7. Sixty respondents (75%) attended at least one RUSSCO contouring workshop. Of those who were aware of eContour, 76% were introduced during a RUSSCO contouring workshop, and 81% continue to use it in their daily practice. The greatest obstacles to using the program were language barrier (51%) and internet access (38%). Nearly 90% reported their contouring practices changed since they started using the program, particularly for delineation of clinical target volumes (57%) and/or organs at risk (46%). More than 97% found the clinical pearls/links to cooperative group protocols in the software helpful in their daily practice. The majority used the contouring program several times per month (43%) or several times per week (41%). Face-to-face contouring instruction in combination with open-access web-based interactive contouring resource had a meaningful impact on perceived quality of radiation oncology contours among Russian practitioners and has the potential to have applications worldwide.


Vertebral compression fractures after stereotactic body radiation therapy: a large, multi-institutional, multinational evaluation.
OBJECTIVE The purpose of this study was to identify factors contributing to an increased risk for vertebral compression fracture (VCF) following stereotactic body radiation therapy (SBRT) for spinal tumors. METHODS A total of 594 tumors were treated with spinal SBRT as primary treatment or re-irradiation at 8 different institutions as part of a multi-institutional research consortium. Patients underwent LINAC-based, image-guided SBRT to a median dose of 20 Gy (range 8-40 Gy) in a median of 1 fraction (range 1-5 fractions). Median patient age was 62 years. Seventy-one percent of tumors were osteolytic, and a preexisting vertebral compression fracture (VCF) was present in 24% of cases. Toxicity was assessed following treatment. Univariate and multivariate analyses were performed using a logistic regression method to determine parameters predictive for post-SBRT VCF. RESULTS At a median follow-up of 10.1 months (range 0.03-57 months), 80% of patients had local tumor control. At the time of last imaging follow-up, at a median of 8.8 months after SBRT, 3% had a new VCF, and 2.7% had a progressive VCF. For development of any (new or progressive) VCF following SBRT, the following factors were predictive for VCF on univariate analysis: short interval from primary diagnosis to SBRT (less than 36.8 days), solitary metastasis, no additional bone metastases, no prior chemotherapy, preexisting VCF, no MRI used for target delineation, tumor volume of 37.3 cm(3) or larger, equivalent 2-Gy-dose (EQD2) tumor of 41.8 Gy or more, and EQD2 spinal cord Dmax of 46.1 Gy or more. Preexisting VCF, solitary metastasis, and prescription dose of 38.4 Gy or more were predictive on multivariate analysis. The following factors were predictive of a new VCF on univariate analysis: solitary metastasis, no additional bone metastases, and no MRI used for target delineation. Presence of a solitary metastasis and lack of MRI for target delineation remained significant on multivariate analysis. CONCLUSIONS A VCF following SBRT is more likely to occur following treatment for a solitary spinal metastasis, reflecting a more aggressive treatment approach in patients with adequately controlled systemic disease. Higher prescription dose and a preexisting VCF also put patients at increased risk for post-SBRT VCF. In these patients, pre-SBRT cement augmentation could be considered to decrease the risk of subsequent VCF.


Automated quantitative multiplex immunofluorescence in situ imaging identifies phospho-S6 and phospho-PRAS40 as predictive protein biomarkers for prostate cancer lethality.
We have witnessed significant progress in gene-based approaches to cancer prognostication, promising early intervention for high-risk patients and avoidance of overtreatment for low-risk patients. However, there has been less advancement in protein-based approaches, even though perturbed protein levels and post-translational modifications are more directly linked with phenotype. Most current, gene expression-based platforms require tissue lysis resulting in loss of structural and molecular information, and hence are blind to tumor heterogeneity and morphological features. Here we report an automated, integrated multiplex immunofluorescence in situ imaging approach that quantitatively measures protein biomarker levels and activity states in defined intact tissue regions where the biomarkers of interest exert their phenotype. Using this approach, we confirm that four previously reported prognostic markers, PTEN, SMAD4, CCND1 and SPP1, can predict lethal outcome of human prostate cancer. Furthermore, we show that two PI3K pathway-regulated protein activities, pS6 (RPS6-phosphoserines 235/236) and pPRAS40 (AKT1S1-phosphothreonine 246), correlate with prostate cancer lethal outcome as well (individual marker hazard ratios of 2.04 and 2.03, respectively). Finally, we incorporate these 2 markers into a novel 5-marker protein signature, SMAD4, CCND1, SPP1, pS6, and pPRAS40, which is highly predictive for prostate cancer-specific death. The ability to substitute PTEN with phospho-markers demonstrates the potential of quantitative protein activity state measurements on intact tissue. In summary, our approach can reproducibly and simultaneously quantify and assess multiple protein levels and functional activities on intact tissue specimens. We believe it is broadly applicable to not only cancer but other diseases, and propose that it should be well suited for prognostication at early stages of pathogenesis where key signaling protein levels and activities are perturbed.


A monolithic immersion metalens for imaging solid-state quantum emitters.
Quantum emitters such as the diamond nitrogen-vacancy (NV) center are the basis for a wide range of quantum technologies. However, refraction and reflections at material interfaces impede photon collection, and the emitters' atomic scale necessitates the use of free space optical measurement setups that prevent packaging of quantum devices. To overcome these limitations, we design and fabricate a metasurface composed of nanoscale diamond pillars that acts as an immersion lens to collect and collimate the emission of an individual NV center. The metalens exhibits a numerical aperture greater than 1.0, enabling efficient fiber-coupling of quantum emitters. This flexible design will lead to the miniaturization of quantum devices in a wide range of host materials and the development of metasurfaces that shape single-photon emission for coupling to optical cavities or route photons based on their quantum state.


[Lymphocytic primary angiitis of the central nervous system with fan-shaped linear enhancement converging to the lateral ventricles: a case report].
We report a case of lymphocytic primary angiitis of the central nervous system (PACNS) with a characteristic gadolinium-enhancement pattern on magnetic resonance imaging (MRI). A 48-year-old, right-handed man presented with a 3-month history of tremor and progressing dementia. Neurologic examination revealed cognitive decline with anterograde amnesia and postural tremor of the fingers. Except for the positive result of serum antinuclear antibody, intense investigations for infectious, rheumatic and neoplastic diseases were negative. Analysis of cerebrospinal fluid showed mild pleocytosis (14 cells/μl). Brain MRI revealed diffuse hyperintense areas in the deep cerebral white matter on T2-weighted images. Gadolinium-enhanced T1-weighted images demonstrated fan-shaped multiple linear enhancements converging to the body of the lateral ventricles. Brain biopsy showed intense infiltration of small lymphocytes without atypia or granuloma mainly within the vessel walls and perivascular spaces. The diagnosis of lymphocytic PACNS was made. Administration of corticosteroid markedly improved the tremor and cognitive dysfunction. MRI after the treatment showed resolution of the abnormal fan-shaped linear enhancement. He returned to his previous occupation. PACNS should be included in the differential diagnosis for fan-shaped linear enhancement converging to the lateral ventricles on MRI in patients with unexplained progressing dementia.


SorghumBase: a web-based portal for sorghum genetic information and community advancement.
SorghumBase provides a community portal that integrates genetic, genomic, and breeding resources for sorghum germplasm improvement. Public research and development in agriculture rely on proper data and resource sharing within stakeholder communities. For plant breeders, agronomists, molecular biologists, geneticists, and bioinformaticians, centralizing desirable data into a user-friendly hub for crop systems is essential for successful collaborations and breakthroughs in germplasm development. Here, we present the SorghumBase web portal ( https://www.sorghumbase.org ), a resource for the sorghum research community. SorghumBase hosts a wide range of sorghum genomic information in a modular framework, built with open-source software, to provide a sustainable platform. This initial release of SorghumBase includes: (1) five sorghum reference genome assemblies in a pan-genome browser; (2) genetic variant information for natural diversity panels and ethyl methanesulfonate (EMS)-induced mutant populations; (3) search interface and integrated views of various data types; (4) links supporting interconnectivity with other repositories including genebank, QTL, and gene expression databases; and (5) a content management system to support access to community news and training materials. SorghumBase offers sorghum investigators improved data collation and access that will facilitate the growth of a robust research community to support genomics-assisted breeding.


Benefit of Prompt Vs Delayed Initiation of Triple Therapy Following an Exacerbation in Patients with COPD in Japan: A Retrospective Cohort Study.
There is currently limited evidence for the optimal timing of triple therapy initiation in Japan, which is crucial for optimizing strategies for the effective treatment of chronic obstructive pulmonary disease (COPD). This study assessed the impact of prompt vs delayed initiation of triple therapy following a COPD exacerbation on clinical and economic outcomes in patients in Japan. Retrospective cohort study of patients in the Medical Data Vision Co., Ltd. database initiating triple therapy as single-inhaler triple therapy (fluticasone furoate/umeclidinium/vilanterol or budesonide/glycopyrronium/formoterol) or multiple-inhaler triple therapy within 180 days of a moderate-to-severe exacerbation (index). For the main analysis, patients were categorized as prompt or delayed initiators, initiating triple therapy within 0-30 days or 31-180 days of index, respectively. Inverse probability of treatment weighting based on propensity scores was used to adjust for measured confounders between prompt and delayed cohorts. For the main analysis, 610 (60.3%) and 402 (39.7%) patients were prompt and delayed initiators, respectively. The rate of subsequent moderate-to-severe exacerbations following index exacerbation was numerically lower in prompt vs delayed initiators (weighted rate ratio 0.95, 95% confidence interval [CI]: 0.74-1.21; P = 0.6603). Time-to-first subsequent moderate-to-severe exacerbation increased significantly in prompt vs delayed initiators (weighted hazard ratio 0.77, 95% CI: 0.64-0.93; P = 0.0053). In patients indexed on a severe exacerbation, delayed initiation resulted in significantly higher 90-day all-cause readmissions vs prompt initiation (42.1% vs 30.6%; P = 0.0329 [weighted estimates]). Weighted healthcare resource utilization rates were numerically lower in prompt vs delayed initiators, and weighted direct costs (all cause and COPD-related) were significantly lower in prompt initiators. This real-world study demonstrated that earlier initiation of triple therapy resulted in several benefits in clinical outcomes for COPD and may also reduce the economic burden of COPD management in Japan.


Sleep Disorders and Chronic Pain Syndromes in the Pediatric Population.
Sleep problems are widespread in children and adolescents suffering from chronic pain disorders. Sleep loss intensifies the experience of pain and is detrimental to the budding self-efficacy of a young individual with limitless horizons. Addressing sleep disorders may prevent the chronification of pain and prevent adverse health outcomes, such as functional impairment, psychiatric comorbidities and overall poor quality of life. This review will explore the cyclical nature between sleep, pain and mood, as well as the functional impact of this relationship on children and adolescents. There will be a discussion about sleep assessment and diagnostic testing, followed by a description of sleep disturbances found in specific pain conditions, ranging from headache, musculoskeletal/abdominal pain, to rheumatologic disorders. Finally, there will be a brief review of pharmacologic and behavioral interventions designed to improve sleep quality, and when possible, to alleviate pain.


Preserving the Efficacy of Glycopeptide Antibiotics during Nanoencapsulation in Liposomes.
Liposome nanovesicles are attractive vehicles for encapsulation and localized delivery of antibiotics. Most liposomal batch preparation processes involve numerous freeze-thaw cycles and heating or sonication steps, all of which can potentially deactivate or degrade antibiotics. We investigated the extent of antibiotic deactivation during various liposomal preparation methods using two glycopeptide antibiotics clinically administered for Staphylococcus infections, namely, vancomycin hydrochloride and teicoplanin. Both antibiotics, in the nonencapsulated state, were found to be highly sensitive to the freeze-thaw/sonication; vancomycin completely lost efficacy after only three cycles of freeze-thaw, and teicoplanin lost efficacy after 20 min of sonication. When the antibiotics were encapsulated in liposomes, vancomycin retained full potency against bacterial cultures of Staphylococcus aureus but encapsulated teicoplanin suffered a decrease in activity. Differential scanning calorimetry and mass spectrometry suggest that liposomes have a protective effect on the encapsulated antibiotic, the extent of which was found to differ on the basis of the processing conditions.


TOLERANCE OF STAPHYLOCOCCUS AUREUS TO SODIUM CHLORIDE.
Parfentjev, I. A. (Institute of Applied Biology, New York, N.Y.), and Anna R. Catelli. Tolerance of Staphylococcus aureus to sodium chloride. J. Bacteriol. 88:1-3. 1964.-The tolerance of Staphylococcus aureus to high concentrations of sodium chloride in liquid medium has been reported. We found that S. aureus grows at 37 C in Tryptose Phosphate Broth saturated with sodium chloride. No difference was noticed between possibly pathogenic and nonpathogenic strains. Under the conditions of our tests, no changes in the original properties of S. aureus strains occurred. In contrast, solutions of sodium chloride in distilled water were injurious to staphylococci and killed most of these organisms in 1 hr. Staphylococci were killed faster at 37 C than at room temperature in a solution of 0.85% sodium chloride in water. Addition of traces of Tryptose Phosphate Broth had a protective effect and prolonged the life of these organisms in physiological saline. All tests were performed at pH 7.2.


[A comparative study of the Feira de Santana (Bahia) and Porto Rico strains of Schistosoma mansoni in experimental infection of mice].
A comparative study of mice infected either with the Porto Rican or a Brazilian (Feira de Santana) strain of Schistosoma mansoni failed to show any difference regarding recovery of five day lung worms, recovery of adult worms from the portal system, number of eggs per gram of tissue in liver and intestines, histopathology or mortality rate. In fact, the differences, although small, were greater among animals infected with the same strain, that when the total results from both strains were compared.


Benzo(a)pyrene-albumin adducts in humans exposed to polycyclic aromatic hydrocarbons in an industrial area of Poland.
The interaction of benzo(a)pyrene with serum albumin was measured in an attempt to identify the actual exposure and to evaluate albumin adduct measurements as biomarkers for exposure monitoring. Benzo(a)pyrene-diol-epoxide (BPDE)-albumin adducts were measured by competitive enzyme linked immunosorbent assay (ELISA) in plasma of coke oven plant workers from three plants and from people living in a highly industrialised area of Silesia in Poland. Due to the high air concentrations of polycyclic aromatic hydrocarbons (PAHs) in this area, a control group was selected from a rural non-industrialised area in Poland. Breathing zone air measurements of PAHs were collected from some of the participants. Coke oven plant workers and non-occupationally exposed people had similar concentrations of albumin adducts whereas the rural controls were significantly lower (2.74 fmol adducts/microgram albumin (SEM 0.124)). The mean concentration of BPDE-albumin adduct in plasma of both the occupational and the environmental groups were significantly higher in the summer samples (4.34 fmol adducts/microgram albumin (SEM 0.335) and 4.55 fmol adducts/microgram albumin (SEM 0.296), respectively) than in the winter samples (3.06 fmol adducts/microgram albumin (SEM 0.187) and 3.04 fmol adducts/microgram albumin (SEM 0.184), respectively) even though the air measurements showed higher concentrations of PAHs in the winter. The statistical analysis did not show any effects of air exposures on concentrations of BPDE-albumin adduct. A multiple regression analysis of the measured concentrations of BPDE-albumin adducts for all the groups, during both seasons, indicates that occupational exposures do not contribute significantly to the formation of adducts. In general, the concentrations of albumin adducts found vary within relatively small limits for the two seasons and between the various groups of participants. No extreme differences were found.


Risk factors for legal induced abortion-related mortality in the United States.
To assess risk factors for legal induced abortion-related deaths. This is a descriptive epidemiologic study of women dying of complications of induced abortions. Numerator data are from the Abortion Mortality Surveillance System. Denominator data are from the Abortion Surveillance System, which monitors the number and characteristics of women who have legal induced abortions in the United States. Risk factors examined include age of the woman, gestational length of pregnancy at the time of termination, race, and procedure. Main outcome measures include crude, adjusted, and risk factor-specific mortality rates. During 1988-1997, the overall death rate for women obtaining legally induced abortions was 0.7 per 100000 legal induced abortions. The risk of death increased exponentially by 38% for each additional week of gestation. Compared with women whose abortions were performed at or before 8 weeks of gestation, women whose abortions were performed in the second trimester were significantly more likely to die of abortion-related causes. The relative risk (unadjusted) of abortion-related mortality was 14.7 at 13-15 weeks of gestation (95% confidence interval [CI] 6.2, 34.7), 29.5 at 16-20 weeks (95% CI 12.9, 67.4), and 76.6 at or after 21 weeks (95% CI 32.5, 180.8). Up to 87% of deaths in women who chose to terminate their pregnancies after 8 weeks of gestation may have been avoidable if these women had accessed abortion services before 8 weeks of gestation. Although primary prevention of unintended pregnancy is optimal, among women who choose to terminate their pregnancies, increased access to surgical and nonsurgical abortion services may increase the proportion of abortions performed at lower-risk, early gestational ages and help further decrease deaths. II-2


Binding and lysing of blood clots using MRX-408.
A thrombus-specific ultrasound contrast agent, MRX-408, has been developed recently. This agent consists of phospholipid-coated microbubbles with a ligand capable of targeting the GPIIb/IIIa receptor, thereby allowing the microbubbles to bind with thrombi rich in activated platelets. In vitro and in vivo animal experiments have been conducted to examine imaging enhancement and sonothrombolysis using this agent compared with a nontargeted agent. For clot binding, blood-smeared slides were incubated with microbubbles and examined under a light microscope. Change in backscatter signals from the blood clots after binding was examined by both an ultrasound scanner and two single-element transducers arranged in a transmitter-receiver pair. For clot lysis, either 1-MHz or 20-KHz ultrasound was used to enhance the lysing effects of MRX-408 with or without urokinase. Evidence of binding was demonstrated under a microscope. In vitro experiments showed that the "acoustic signature", or properties, of blood clots changed after binding. Clots became more echogenic and nonlinear. In vivo fundamental ultrasound imaging confirmed that as a result of binding, blood clots were more visible, the area of detection was improved, and shadowing behind clots was more noticeable. Under 1-MHz ultrasound and 30 minutes of treatment, lysis efficiency reached 34% with MRX-408, whereas there was no visible clot lysis with saline. The results of these preliminary studies show that as a contrast agent, MRX-408 enhanced clots under ultrasound imaging and facilitated sonothrombolysis with or without thrombolytic drugs.


The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells.
This study investigated the influence of expression of proteins of the INK4 family, particularly p16, on the growth and self-renewal kinetics of hematopoietic cells. First, retrovirus-mediated gene transfer (RMGT) was used to restore p16(INK4a) expression in the p16(INK4a)-deficient lymphoid and myeloid cell lines BV173 and K562, and it was confirmed that this inhibited their growth. Second, to sequester p16(INK4a) and related INK4 proteins, cyclin-dependent kinase 4 (CDK4) was retrovirally transduced into normal human CD34(+) bone marrow cells and then cultured in myeloid colony-forming cell (CFC) assays. The growth of CDK4-transduced colonies was more rapid; the cell-doubling time was reduced; and, upon replating, the colonies produced greater yields of secondary colonies than mock-untransduced controls. Third, colony formation was compared by marrow cells from p16(INK4a-/-) mice and wild-type mice. The results from p16(INK4a-/-) marrow were similar to those from CDK4-transduced human CFCs, in terms of growth rate and replating ability, and were partially reversed by RMGT of p16(INK4a). Lines of immature granulocytic cells were raised from 15 individual colonies grown from the marrow of p16(INK4a-/-) mice. These had a high colony-forming ability (15%) and replating efficiency (96.7%). The p16(INK4a-/-) cell lines readily became growth factor-independent upon cytokine deprivation. Taken together, these results demonstrate that loss of INK4 proteins, in particular p16(INK4a), increases the growth rate of myeloid colonies in vitro and, more importantly, confers an increased ability for clonal expansion on hematopoietic progenitor cells.