Cryptic population structuring and the role of the Isthmus of Tehuantepec as a gene flow barrier in the critically endangered Central American River Turtle.
The critically endangered Central American River Turtle (Dermatemys mawii) is the only remaining member of the Dermatemydidae family, yet little is known about its population structuring. In a previous study of mitochondrial (mt) DNA in the species, three main lineages were described. One lineage (Central) was dominant across most of the range, while two other lineages were restricted to Papaloapan (PAP; isolated by the Isthmus of Tehuantepec and the Sierra de Santa Marta) or the south-eastern part of the range (1D). Here we provide data from seven polymorphic microsatellite loci and the R35 intron to re-evaluate these findings using DNA from the nuclear genome. Based on a slightly expanded data set of a total of 253 samples from the same localities, we find that mtDNA and nuclear DNA markers yield a highly congruent picture of the evolutionary history and population structuring of D. mawii. While resolution provided by the R35 intron (sequenced for a subset of the samples) was very limited, the microsatellite data revealed pronounced population structuring. Within the Grijalva-Usumacinta drainage basin, however, many populations separated by more than 300 kilometers showed signals of high gene flow. Across the entire range, neither mitochondrial nor nuclear DNA show a significant isolation-by-distance pattern, but both genomes highlight that the D. mawii population in the Papaloapan basin is genetically distinctive. Further, both marker systems detect unique genomic signals in four individuals with mtDNA clade 1D sampled on the southeast edge of the Grijalva-Usumacinta basin. These individuals may represent a separate cryptic taxon that is likely impacted by recent admixture.


The use of magnetic resonance spectroscopy in the subacute evaluation of athletes recovering from single and multiple mild traumatic brain injury.
Advanced neuroimaging techniques have shown promise in highlighting the subtle changes and nuances in mild traumatic brain injury (MTBI) even though clinical assessment has shown a return to pre-injury levels. Here we use ¹H-magnetic resonance spectroscopy (¹H-MRS) to evaluate the brain metabolites N-acetyl aspartate (NAA), choline (Cho), and creatine (Cr) in the corpus callosum in MTBI. Specifically, we looked at the NAA/Cho, NAA/Cr, and Cho/Cr ratios in the genu and splenium. We recruited 20 normal volunteers (NV) and 28 student athletes recovering from the subacute phase of MTBI. The MTBI group was categorized based upon the number of MTBIs and time from injury to ¹H-MRS evaluation. Significant reductions in NAA/Cho and NAA/Cr ratios were seen in the genu of the corpus callosum, but not in the splenium, for MTBI subjects, regardless of the number of MTBIs. MTBI subjects recovering from their first MTBI showed the greatest alteration in NAA/Cho and NAA/Cr ratios. Time since injury to ¹H-MRS acquisition was based upon symptom resolution and did not turn out to be a significant factor. We observed that as the number of MTBIs increased, so did the length of time for symptom resolution. Unexpected findings from this study are that MTBI subjects showed a trend of increasing NAA/Cho and NAA/Cr ratios that coincided with increasing number of MTBIs.


Network Analysis of Protein Adaptation: Modeling the Functional Impact of Multiple Mutations.
The evolution of new biochemical activities frequently involves complex dependencies between mutations and rapid evolutionary radiation. Mutation co-occurrence and covariation have previously been used to identify compensating mutations that are the result of physical contacts and preserve protein function and fold. Here, we model pairwise functional dependencies and higher order interactions that enable evolution of new protein functions. We use a network model to find complex dependencies between mutations resulting from evolutionary trade-offs and pleiotropic effects. We present a method to construct these networks and to identify functionally interacting mutations in both extant and reconstructed ancestral sequences (Network Analysis of Protein Adaptation). The time ordering of mutations can be incorporated into the networks through phylogenetic reconstruction. We apply NAPA to three distantly homologous β-lactamase protein clusters (TEM, CTX-M-3, and OXA-51), each of which has experienced recent evolutionary radiation under substantially different selective pressures. By analyzing the network properties of each protein cluster, we identify key adaptive mutations, positive pairwise interactions, different adaptive solutions to the same selective pressure, and complex evolutionary trajectories likely to increase protein fitness. We also present evidence that incorporating information from phylogenetic reconstruction and ancestral sequence inference can reduce the number of spurious links in the network, whereas preserving overall network community structure. The analysis does not require structural or biochemical data. In contrast to function-preserving mutation dependencies, which are frequently from structural contacts, gain-of-function mutation dependencies are most commonly between residues distal in protein structure.


In vitro and in vivo investigation of low molecular weight heparin-alginate beads for oral administration.
Low molecular weight heparin (LMWH) and standard heparin are widely used anticoagulants. However, they have very poor oral bioavailability and have to be administered by the parenteral route. Alginates are biodegradable, biocompatible and mucoadhesive polymers which can be used for advantage for the oral administration of LMWH. The aim of the study was to develop LMWH-alginate beads for oral delivery. Alginate beads were prepared based on the 2(3) factorial design. In vitro characterization studies of the beads were carried out. In vivo studies were performed on rabbits. The LMWH solutions (5000 IU/kg, with and without 5% dimethyl-β-cyclodextrin), as well as the LMWH-alginate beads were administered to rabbits. The IV solution was also administered (100 IU/kg). The anti-Xa activity was measured in plasma. Area under curve (AUC) and Cmax values were determined. Histological investigations were also carried out. The formulation consisting of a 1:2 drug/alginate ratio and cured using 0.5 M CaCl2 for 15 min gave the best result in terms of encapsulation efficiency and the time for 50% of the drug to be released (t50%). A significantly higher bioavailability was observed for LMWH-alginate beads than for LMWH solutions. It was concluded that, anticoagulant effectiveness was achieved using alginate beads containing LMWH after oral administration to the rabbits.


Postexposure treatment with doxycycline for the prevention of tick-borne relapsing fever.
Tick-borne relapsing fever (TBRF) is an acute febrile illness. In Israel, TBRF is caused by Borrelia persica and is transmitted by Ornithodoros tholozani ticks. We examined the safety and efficacy of postexposure treatment to prevent TBRF. In a double-blind, placebo-controlled trial, 93 healthy subjects with suspected tick exposure (52 with signs of tick bites and 41 close contacts--those without signs but with a similar risk of contact with ticks) were randomly assigned to receive either doxycycline (Dexxon, in a dose of 200 mg the first day and then 100 mg per day for four days) or placebo after presumed exposure to TBRF. Cases of TBRF were defined by fever and a positive blood smear. Serologic analysis for cross-reactivity to Borrelia burgdorferi and polymerase chain reaction (PCR) for the borrelia glpQ gene were also performed. After randomization, 47 subjects (26 with signs of tick bites and 21 close contacts) received doxycycline. Forty-six other subjects (26 with signs of tick bites and 20 close contacts) received placebo. All 10 cases of TBRF identified by a positive blood smear were in the placebo group of subjects with signs of a tick bite (P<0.001). These findings suggested a 100 percent efficacy of preemptive treatment (95 percent confidence interval, 46 to 100 percent). PCR for the borrelia glpQ gene was negative at baseline for all subjects and subsequently positive in all subjects with fever and a positive blood smear. Seroconversion was detected in eight of nine cases of TBRF. PCR and serum samples were negative for all of the other subjects tested. No major treatment-associated adverse effects were identified. Treatment with doxycycline is safe and efficacious in preventing TBRF after suspected exposure to ticks in a high-risk environment. (ClinicalTrials.gov number, NCT00237016 [ClinicalTrials.gov].).


Participant referral rate in the National Eye Health Survey (NEHS).
To present the rates of referral of participants in the National Eye Health Survey (NEHS) for further eye care. A national sample of 3098 non-Indigenous Australians aged 50-98 and 1738 Indigenous Australians aged 40-92 years living in 30 randomly selected sites was recruited using a door-to-door approach. Participants completed a general questionnaire and a series of eye tests, including vision and anterior segment assessment, intra-ocular pressure measurement, visual field testing and fundus photography. A predefined protocol was used to guide the referral of participants for follow up eye care. An ophthalmologist was on-call to assist with the triaging of participants. Of the total sample, 32.1% (994/3098) of non-Indigenous participants and 43.6% (757/1738) of Indigenous participants were referred for further eye care (p<0.001). A significant difference in referral rates for Indigenous Australians was observed between regions of differing geographic remoteness [range = 32.2% (Inner Regional)-60.4% (Very Remote), p <0.001]. After adjusting for covariates, males (OR = 1.24, 95% CI: 1.06-1.46), older age (OR = 1.02 per year, 95% CI: 1.01, 1.02) and longer time since previous eye examination (OR = 1.15 per year, 95% CI: 1.12, 1.19) were associated with higher rates of eye care referral in the non-Indigenous population. In the Indigenous population, older age (OR = 1.02 per year, 95% CI: 1.01-1.03), self-reported diabetes (OR = 1.70, 95% CI: 1.37-2.12), greater geographical remoteness (OR = 1.19, 95% CI: 1.09-1.29) and longer time since previous eye examination (OR = 1.10 per year, 95% CI: 1.07, 1.13) were associated with a higher rate of referral after multivariate adjustments. A total of 25 participants (1.4%) were referred for urgent follow-up of potentially sight threatening conditions. Our data has identified several high risk groups that required ophthalmic referral including older Australians, non-Indigenous men, Indigenous Australians with self-reported diabetes and those residing in very remote populations who may benefit from improvements in the provision and/or uptake of eye health services. Future longitudinal research is warranted to evaluate the feasibility and efficacy of implementing a referral protocol within a population-based research setting.


Within-farm transmission characteristics of bluetongue virus serotype 8 in cattle and sheep in the Netherlands, 2007-2008.
In 2006 and 2007, sheep and cattle farms in the Netherlands were affected by an epidemic of bluetongue virus serotype 8 (BTV-8). In order to obtain insight into the within-farm spread of the virus, five affected cattle and five affected sheep farms were longitudinally monitored between early 2007 and mid or late 2008. The farms were visited between four and seven times to collect blood samples. During each visit, all animals present in the flock or herd were sampled. The samples were analysed for the presence of BTV-8 antibodies (ELISA) and BTV-8 antigen (rRT-PCR). The observed patterns of RT-PCR positives indicate a rapid within-farm virus spread during the vector season. During vector-free periods we observed a complete rRT-PCR positivity decline within a few months. During the vector season a lower bound estimate of the basic reproduction number (R0) ranges from 2.9-6.9 in the cattle herds (one herd not analysed), and from 1.3-3.2 in the sheep flocks in this study.


[Food consumption: in search of a multidisciplinary approach].
Changes in eating habits have been observed in many countries in recent decades revealing the complexity of consumption models and their determining factors. Thus it is that the adoption of a multidisciplinary and comparative approach by which the various aspects of eating habits (economic, social, cultural and nutritional) may be assessed so as to permit the elucidation of the mechanisms responsible for these changes and their consequences in different socio-economic contexts, becomes even more necessary. The contributions of the various areas of knowledge to the study of the nutritional situation are here analysed and the approach to this theme in countries of the North and the South is discussed, as is, specifically, the situation in Brazil. Finally, the analysis of the consumption of food-stuffs on the basis of the notion of "food systems", within a historical perspective, is proposed. The various social agents (producers, distributors, consumers, State), their systems of logic, their strategies and the relationships which they establish among themselves over time are taken into account in such a way as to make possible an understanding of the process by which eating habits are constructed and developed.


Subclinical atherosclerosis in patients with rheumatoid arthritis. A meta-analysis of literature studies.
We performed a systematic review with meta-analysis and meta-regression of literature studies evaluating the impact of rheumatoid arthritis (RA) on common carotid artery intima-media thickness (CCA-IMT) and on the prevalence of carotid plaques. Studies evaluating the relationship between RA and markers of cardiovascular (CV) risk (CCA-IMT and prevalence of carotid plaques) were systematically searched in the PubMed, Web of Science, Scopus and EMBASE databases. A total of 59 studies (4,317 RA patients and 3,606 controls) were included in the final analysis, 51 studies with data on CCA-IMT (52 data-sets on 3,600 RA patients and 3,020 controls) and 35 studies reporting on the prevalence of carotid plaques (2,859 RA patients and 2,303 controls). As compared to controls, RA patients showed a higher CCA-IMT (mean difference [MD]: 0.10 mm; 95 % confidence interval [CI]: 0.07, 0.12; p < 0.00001), and an increased prevalence of carotid plaques (odds ratio [OR]: 3.61; 95 %CI: 2.65, 4.93; p< 0.00001). Interestingly, when analysing studies on early RA, the difference in CCA-IMT among RA patients and controls was even higher (MD: 0.21 mm; 95 %CI: 0.06, 0.35; p=0.006), and difference in the prevalence of carotid plaques was entirely confirmed (OR: 3.57; 95 %CI: 1.69, 7.51; p=0.0008). Meta-regression models showed that male gender and a more severe inflammatory status [as expressed by disease activity score in 28 joints (DAS28), C-reactive protein (CRP) levels, and erythrocyte sedimentation rate (ESR)] significantly impacted on CCA-IMT. In conclusion, RA appears significantly associated with subclinical atherosclerosis and CV risk. These findings can be useful to plan adequate prevention strategies and therapeutic approaches.


[Clinical and functional changes in the biliary tract in patients with psoriasis concurrent with chronic opisthorchiasis].
to study the clinical and functional changes in the biliary tract in patents with psoriasis concurrent with chronic opisthorchiasis (CO). 60 patients with psoriasis concurrent with CO, 45 patients with psoriasis without helminthism, 30 with CO, and 15 healthy individuals were examined. Chromatic fractional duodenal intubation, dynamic hepatobiliscintigraphy, and ultrasound study of the liver and gallbladder were used. The concentrations of cholesterol, bile acids, bilirubin, total lipids and phospholipids in bile were measured. A number of relative indices, such as cholate-cholesterol coefficient (CCC), Thomas-Hofmann index, and Rubens index were calculated. The patients with psoriasis with CO were found to have signs of mainly hypermotor dyskinesia of the biliary tract, as well as increased bile lithogenicity (lower CCC, and increases in the Rubens index and Thomas-Hofmann index). The clinical and functional changes found in the biliary tract of the patients with psoriasis concurrent with CO show it necessary to perform antihelminthic therapy in this group of patients.


A descriptive study of US Special Operations Command fatalities, 2001 to 2018.
Studies of fatalities from injury and disease guide prevention and treatment efforts for populations at risk. Findings can inform leadership and direct clinical practice guidelines, research, and personnel, training, and equipment requirements. A retrospective review and descriptive analysis was conducted of United States Special Operations Command (USSOCOM) fatalities who died while performing duties from September 11, 2001, to September 10, 2018. Characteristics analyzed included subcommand, military activity, operational posture, and manner of death. Of 614 USSOCOM fatalities (median age, 30 years; male, 98.5%) the leading cause of death was injury (97.7%); specifically, multiple/blunt force injury (34.5%), blast injury (30.7%), gunshot wound (GSW; 30.3%), and other (4.5%). Most died outside the United States (87.1%), during combat operations (85.3%), in the prehospital environment (91.5%), and the same day of insult (90.4%). Most fatalities were with the US Army Special Operations Command (67.6%), followed by the Naval Special Warfare Command (16.0%), Air Force Special Operations Command (9.3%), and Marine Corps Forces Special Operations Command (7.2%). Of 54.6% who died of injuries incurred during mounted operations, most were on ground vehicles (53.7%), followed by rotary-wing (37.3%) and fixed-wing (9.0%) aircrafts. The manner of death was primarily homicide (66.0%) and accident (30.5%), followed by natural (2.1%), suicide (0.8%), and undetermined (0.7%). Specific homicide causes of death were GSW (43.7%), blast injury (42.2%), multiple/blunt force injury (13.8%), and other (0.2%). Specific accident causes of death were multiple/blunt force injury (80.7%), blast injury (6.4%), GSW (0.5%), and other (12.3%). Of accident fatalities with multiple/blunt force injury, the mechanism was mostly aircraft mishaps (62.9%), particularly rotary wing (68.4%). Most USSOCOM fatalities died abroad from injury in the prehospital setting. To improve survival from military activities worldwide, leaders must continue to optimize prehospital capability and develop strategies that rapidly connect patients to advanced resuscitative and surgical care. Epidemiological, level IV; Therapeutic level IV.


Factors associated with antibiotic prescribing for acute bronchitis at a university health center.
Antibiotics are not indicated for treating acute bronchitis cases, yet up to 70% of adult acute bronchitis medical visits in the USA result in an antibiotic prescription. Reducing unnecessary antibiotic prescribing for acute bronchitis is a key antibiotic stewardship goal set forth by the Centers for Disease Control and Prevention. Understanding what factors influence prescribing for bronchitis cases can inform antimicrobial stewardship initiatives. The goal of this study was to identify factors associated with antibiotic prescribing at a high-volume student health center at a large US university. The Pennsylvania State University Health Services offers on-campus medical care to a population of over 40,000 students and receives over 50,000 visits every year. We conducted a retrospective chart review of acute bronchitis visits for the 2015-2016 academic year and used a multivariate logistic regression analysis to identify variables associated with antibiotic prescribing. Findings during lung exams increased the likelihood of an antibiotic prescription (rales OR 13.95, 95% CI 3.31-80.73; rhonchi OR 5.50, 95% CI 3.08-10.00; percussion abnormality OR 13.02, 95% CI 4.00-50.09). Individual clinicians had dramatically different rates of prescribing (OR range 0.03-12.3). Male patients were more likely than female patients to be prescribed antibiotics (OR 1.68, 95% CI 1.17-2.41). Patients who reported longer duration since the onset of symptoms were slightly more likely to receive prescriptions (OR 1.04 per day, 95% CI 1.03-1.06), as were patients who reported worsening symptoms (OR 1.78, 95% CI 1.03-3.10). Visits with diagnoses or symptoms associated with viral infections or allergies were less likely to result in prescriptions (upper respiratory tract infection (URI) diagnosis OR 0.33, 95% CI 0.18-0.58; sneezing OR 0.39, 95% CI 0.17-0.86; vomiting OR 0.31, 95% CI 0.10-0.83). An exam finding of anterior cervical lymphadenopathy was associated with antibiotic prescribing (tender OR 3.85, 95% CI 1.70-8.83; general OR 2.63, 95% CI 1.25-5.54). Suspicious findings during lung examinations (rales, rhonchi, percussion abnormality) and individual healthcare providers were important factors influencing antibiotic prescribing rates for acute bronchitis visits. Patient gender, worsening symptoms, duration of illness, symptoms associated with viral infections or allergies, and anterior cervical lymphadenopathy also influenced prescribing rates.


Pulmonary Hypertension by the Method of Paul Wood.
A physiological approach to the analysis of hemodynamic data in pulmonary hypertension (PH) has the advantage of reducing the large number (well over 100) of potential causal illnesses into four simple mechanisms. A fifth condition is composed of mixtures of the four basic mechanisms. This approach was beautifully described by Paul Wood, the great cardiologist whose name is given to the units of pulmonary vascular resistance (PVR), Wood units. This approach uses well understood physiological contributions to pulmonary vascular pressure. It is powerful, the major uncertainty being in determination of the magnitude of each mechanism in patients that have mixed PH of several causes. It also makes sense of the occasionally awkward clustering of conditions in the clinical classification of the World Symposium, which omits pulmonary vasoconstriction, hyperkinetic states, and the highly prevalent condition of "mixed" PH. This method of analysis is described and demonstrated, much as Wood did in his writings. The method is useful in the office, the ICU, and in consultation. A basic message from this approach is that correct assessment requires measurement of each of the three major inputs, pulmonary arterial pressure (Ppa), pulmonary artery wedge pressure (Pwedge) and cardiac output (CO). Some cases also need left ventricular end diastolic pressure (LVEDP). Other data contributing to analysis will be discussed in each condition. A key to avoiding mistakes is to always remember that PH is simply an elevation in pressure and is not inherently diagnostic of cause.


Selective packaging of human growth hormone into synaptic vesicles in a rat neuronal (PC12) cell line.
We have introduced the gene for human growth hormone (hGH) into PC12 cells, a rat pheochromocytoma-derived cell line with neuronal characteristics, and have isolated stable cell lines that express this protein. hGH is stored within the cells in membrane-bounded vesicles that are indistinguishable from the endogenous catecholaminergic synaptic vesicles. When the transfected cells are stimulated by carbachol or direct depolarization, they release norepinephrine and hGH with parallel kinetics. Treatment of the transfected cells with nerve growth factor results in a twofold increase in the amounts of hGH stored in and secreted from the cells. Not all proteins are packaged into the synaptic vesicles, since the rate of release of laminin, a soluble secreted protein endogenous to PC12 cells, is not stimulated by carbachol. This neuronal cell line therefore possesses at least two distinct pathways for secretion and can selectively package a foreign endocrine hormone into the regulated pathway.


What happened? Alcohol, memory blackouts, and the brain.
Alcohol primarily interferes with the ability to form new long-term memories, leaving intact previously established long-term memories and the ability to keep new information active in memory for brief periods. As the amount of alcohol consumed increases, so does the magnitude of the memory impairments. Large amounts of alcohol, particularly if consumed rapidly, can produce partial (i.e., fragmentary) or complete (i.e., en bloc) blackouts, which are periods of memory loss for events that transpired while a person was drinking. Blackouts are much more common among social drinkers--including college drinkers--than was previously assumed, and have been found to encompass events ranging from conversations to intercourse. Mechanisms underlying alcohol-induced memory impairments include disruption of activity in the hippocampus, a brain region that plays a central role in the formation of new autobiographical memories.


A novel technique for positioning multiple cell types by liquid handling.
The spatial control of cells on a surface and the patterning of multiple cell types is an important tool for fundamental biological research and tissue engineering applications. A novel technique is described for the controlled seeding of multiple cell types at specific locations on a surface without requiring the use of specialized equipment or materials. Small-volume, quasi-hemispherical drops of cell solution are deposited onto a cell culture surface immersed under barrier oil, which serves to contain the drop and prevents evaporation of the cell culture medium during the time necessary for cells to attach to the cell culture surface. Subsequent flooding with an aqueous cell-compatible buffer displaces the barrier oil, allowing the cells to grow freely across the surface. This technique offers a simple and easily implemented solution for defining the initial position of cultured cells. The coculture of multiple cell types may be carried out by incorporating different cell types in each drop. A suitable drop volume was found to be 1 microl dispensed with a standard 0.5-10 microl pipette. The drop formed resulted in a footprint diameter of approximately 2 mm. Mineral oil and silicone oil do not compromise the viability of cultured cells when used in this technique. Moreover, a surface with heparin-immobilized FGF2 is shown to retain its bioactivity following drying of the substrate and contact with mineral oil.


Prevalence, risk factors and molecular characterization of Chlamydia trachomatis in pregnant women from Córdoba, Argentina: A prospective study.
Chlamydia trachomatis causes the most prevalent bacterial Sexual Transmitted Infection. In pregnant women, untreated chlamydial infections are associated with abortions, premature rupture of membranes, postpartum endometritis, low birth weight and transmission to the newborn. In Córdoba, Argentina, there is little knowledge about the prevalence of Chlamydia trachomatis in women in their third trimester of pregnancy, so, the aim of this study was to evaluate Chlamydia trachomatis prevalence and genotypes present in Cordovan pregnant women with different age and socioeconomic status. Design: prospective study. Settings: Women population from Cordoba city, Argentina. Population: Pregnant women having 35 to 37 weeks of gestation. Methods: Five hundred and nine cervical swabs were collected. Each sample was subjected to DNA extraction and PCR for Chlamydia trachomatis using primers NRO/NLO and CTP1/CTP2. Positives samples were sequenced to determine genotype. Main outcome measures: Demographic data of the patients were collected to detect a population at risk for this infection. A prevalence of 6.9% (35/509) for Chlamydia trachomatis infection was detected, with 32/295 and 3/214 from pregnant women with low or better economic resources respectively (p = 0,0001). Results showed a significantly increased rate of 11.6% (30/258) in women under 25 years compared with 2% (5/251) in patients over that age (p = 0,00003). Genotype E was the most prevalent. With these results, we can say that pregnant women under 25 years old and low economic resources are one of the populations in which the screening programs of Chlamydia trachomatis should focus.


A Shift in ApoM/S1P Between HDL-Particles in Women With Type 1 Diabetes Mellitus Is Associated With Impaired Anti-Inflammatory Effects of the ApoM/S1P Complex.
Type 1 diabetes mellitus (T1D) patients have an increased risk of cardiovascular disease despite high levels of high-density lipoproteins (HDL). Apolipoprotein M (apoM) and its ligand sphingosine 1-phospate (S1P) exert many of the anti-inflammatory effects of HDL. We investigated whether apoM and S1P are altered in T1D and whether apoM and S1P are important for HDL functionality in T1D. ApoM and S1P were quantified in plasma from 42 healthy controls and 89 T1D patients. HDL was isolated from plasma and separated into dense, medium-dense, and light HDL by ultracentrifugation. Primary human aortic endothelial cells were challenged with tumor necrosis factor-α in the presence or absence of isolated HDL. Proinflammatory adhesion molecules E-selectin and vascular cellular adhesion molecule-1 were quantified by flow cytometry. Activation of the S1P1- receptor was evaluated by analyzing downstream signaling targets and receptor internalization. There were no differences in plasma levels of apoM and S1P between controls and T1D patients, but the apoM/S1P complexes were shifted from dense to light HDL particles in T1D. ApoM/S1P in light HDL particles from women were less efficient in inhibiting expression of vascular cellular adhesion molecule-1 than apoM/S1P in denser particles. The light HDL particles were unable to activate Akt, whereas all HDL subfractions were equally efficient in activating Erk and receptor internalization. ApoM/S1P in light HDL particles were inefficient in inhibiting tumor necrosis factor-α-induced vascular cellular adhesion molecule-1 expression in contrast to apoM/S1P in denser HDL particles. T1D patients have a higher proportion of light particles and hence more dysfunctional HDL, which could contribute to the increased cardiovascular disease risk associated with T1D.


The importance of biotic interactions and local adaptation for plant response to environmental changes: field evidence along an elevational gradient.
Predicting the response of species to environmental changes is a great and on-going challenge for ecologists, and this requires a more in-depth understanding of the importance of biotic interactions and the population structuration in the landscape. Using a reciprocal transplantation experiment, we tested the response of five species to an elevational gradient. This was combined to a neighbour removal treatment to test the importance of local adaptation and biotic interactions. The trait studied was performance measured as survival and biomass. Species response varied along the elevational gradient, but with no consistent pattern. Performance of species was influenced by environmental conditions occurring locally at each site, as well as by positive or negative effects of the surrounding vegetation. Indeed, we observed a shift from competition for biomass to facilitation for survival as a response to the increase in environmental stress occurring in the different sites. Unlike previous studies pointing out an increase of stress along the elevation gradient, our results supported a stress gradient related to water availability, which was not strictly parallel to the elevational gradient. For three of our species, we observed a greater biomass production for the population coming from the site where the species was dominant (central population) compared to population sampled at the limit of the distribution (marginal population). Nevertheless, we did not observe any pattern of local adaptation that could indicate adaptation of populations to a particular habitat. Altogether, our results highlighted the great ability of plant species to cope with environmental changes, with no local adaptation and great variability in response to local conditions. Our study confirms the importance of taking into account biotic interactions and population structure occurring at local scale in the prediction of communities' responses to global environmental changes.


Anaerobic bacteria as a delivery system for cancer gene therapy: in vitro activation of 5-fluorocytosine by genetically engineered clostridia.
Certain species of anaerobic bacteria have been shown to localise and germinate specifically in the hypoxic regions of tumours, resulting in tumour lysis. We propose an innovative approach to cancer gene therapy in which genetically engineered anaerobic bacteria of the genus Clostridium are used to achieve tumour-specific gene delivery. Our strategy involves enzyme/prodrug therapy, in which the Escherichia coli enzyme cytosine deaminase is used to convert the non-toxic prodrug 5-fluorocytosine to the active chemotherapeutic agent 5-fluorouracil. The E. coli gene encoding cytosine deaminase has been cloned into a clostridial expression vector and transformed into Clostridium beijerinckii, resulting in constitutive expression of cytosine deaminase and significant levels of active enzyme in the bacterial medium. When added to an in vitro clonogenic survival assay, supernatant from clostridia expressing cytosine deaminase increased the sensitivity of murine EMT6 carcinoma cells to 5-fluorocytosine approximately 500-fold. This high level of prodrug activation, combined with the specificity of clostridia for hypoxic regions of tumours, indicates a potential use in cancer gene therapy.


Social Support and Common Dyadic Coping in Couples' Dyadic Management of Type II Diabetes: Protocol for an Ambulatory Assessment Application.
Type II diabetes mellitus (T2DM) is a common chronic disease. To manage blood glucose levels, patients need to follow medical recommendations for healthy eating, physical activity, and medication adherence in their everyday life. Illness management is mainly shared with partners and involves social support and common dyadic coping (CDC). Social support and CDC have been identified as having implications for people's health behavior and well-being. Visible support, however, may also be negatively related to people's well-being. Thus, the concept of invisible support was introduced. It is unknown which of these concepts (ie, visible support, invisible support, and CDC) displays the most beneficial associations with health behavior and well-being when considered together in the context of illness management in couple's everyday life. Therefore, a novel ambulatory assessment application for the open-source behavioral intervention platform MobileCoach (AAMC) was developed. It uses objective sensor data in combination with self-reports in couple's everyday life. The aim of this paper is to describe the design of the Dyadic Management of Diabetes (DyMand) study, funded by the Swiss National Science Foundation (CR12I1_166348/1). The study was approved by the cantonal ethics committee of the Canton of Zurich, Switzerland (Req-2017_00430). This study follows an intensive longitudinal design with 2 phases of data collection. The first phase is a naturalistic observation phase of couples' conversations in combination with experience sampling in their daily lives, with plans to follow 180 T2DM patients and their partners using sensor data from smartwatches, mobile phones, and accelerometers for 7 consecutive days. The second phase is an observational study in the laboratory, where couples discuss topics related to their diabetes management. The second phase complements the first phase by focusing on the assessment of a full discussion about diabetes-related concerns. Participants are heterosexual couples with 1 partner having a diagnosis of T2DM. The AAMC was designed and built until the end of 2018 and internally tested in March 2019. In May 2019, the enrollment of the pilot phase began. The data collection of the DyMand study will begin in September 2019, and analysis and presentation of results will be available in 2021. For further research and practice, it is crucial to identify the impact of social support and CDC on couples' dyadic management of T2DM and their well-being in daily life. Using AAMC will make a key contribution with regard to objective operationalizations of visible and invisible support, CDC, physical activity, and well-being. Findings will provide a sound basis for theory- and evidence-based development of dyadic interventions to change health behavior in the context of couple's dyadic illness management. Challenges to this multimodal sensor approach and its feasibility aspects are discussed. PRR1-10.2196/13685.


Index of biventricular interdependence calculated using cardiac MRI: a proof of concept study in patients with and without constrictive pericarditis.
We sought to propose a magnetic resonance (MR) imaging-derived index of biventricular interdependence as a diagnostic parameter to distinguish patients with surgically-confirmed pericardial constriction from those without. Free-breathing real time MR pulse sequences of seventeen subjects with surgically proven constrictive pericarditis and thirty-five patients referred for clinically-indicated cardiac MR examinations but without documented constriction were analyzed using a novel index of biventricular interdependence. Cross-sectional biventricular areas at end diastole using the epicardial surface were traced at the mid left ventricular level at end-inspiration and end-expiration and an index of biventricular interdependence, defined as the ratio of (biventricular end-diastolic area at end-inspiration)/(biventricular end-diastolic area at end-expiration) was calculated for each subject. The mean index for both groups was calculated and results were statistically compared. The index of biventricular interdependence approximated unity (mean index 1.03 ± 0.03 SD) in patients with surgically confirmed pericardial constriction, indicating similar biventricular area at end-inspiration and end-expiration, and was significantly lower than in individuals without constrictive pericarditis (mean index 1.28 ± 0.10 SD; p < 0.0001). The MR-derived index of biventricular interdependence was significantly different between subjects with surgically-confirmed pericardial constriction and subjects where pericardial constraint was not suspected and may serve as a useful metric in the hemodynamic assessment of patients with a potential diagnosis of constrictive pericarditis.


A randomized controlled trial of a community-based dementia care coordination intervention: effects of MIND at Home on caregiver outcomes.
To assess whether MIND at Home, a community-based, multicomponent, care coordination intervention, reduces unmet caregiving needs and burden in informal caregivers of persons with memory disorders. An 18-month randomized controlled trial of 289 community-living care recipient (CR)-caregiver (informal caregivers, i.e., unpaid individuals who regularly assisted the CR) dyads from 28 postal code areas of Baltimore, Maryland was conducted. All dyads and the CR's primary care physician received the written needs assessment results and intervention recommendations. Intervention dyads then received an 18-month care coordination intervention delivered by nonclinical community workers to address unmet care needs through individualized care planning, referral and linkage to dementia services, provision of caregiver dementia education and skill-building strategies, and care progress monitoring by an interdisciplinary team. Primary outcome was total percent of unmet caregiver needs at 18 months. Secondary outcomes included objective and subjective caregiver burden measures, quality of life (QOL), and depression. Total percent of unmet caregiver needs declined in both groups from baseline to 18 months, with no statistically significant between-group difference. No significant group differences occurred in most caregiver burden measures, depression, or QOL. There was a potentially clinically relevant reduction in self-reported number of hours caregivers spent with the CR for MIND participants compared with control subjects. No statistically significant impacts on caregiver outcomes were found after multiple comparison adjustments. However, MIND at Home appeared to have had a modest and clinically meaningful impact on informal caregiver time spent with CRs.


Comparison of pre-contact joint kinematics and vertical impulse between vertical jump landings and step-off landings from equal heights.
Although impact phase differences between vertical jump landings (VJL) and step-off landings (STL) may be related to task-specific pre-contact strategies, pre-contact mechanics are rarely examined. Thus, pre-contact kinematics and vertical ground reaction force (vGRF) impulse were examined between VJL and STL. Ten health adults (20.9 ± 1.6 yrs; 167.8 ± 4.2 cm; 68.5 ± 7.15 kg) performed 15 VJL and 15 STL from equal heights. Limb (lead; trail) by task (VJL; STL) ANOVAs (α = 0.05) compared hip, knee, and ankle joint angles 150 ms pre-contact, 100 ms pre-contact, 50 ms pre-contact, and at ground contact. Joint angular displacement was also evaluated between 150 ms pre-contact and ground contact. vGRF impulse was compared during the loading (ground contact to peak vGRF) and attenuation (peak vGRF to end of impact) phases. Greater hip flexion angles occurred during STL versus VJL at each event except 150 ms pre-contact (p ≤ .004). Trail limb knee flexion angles were greater at each event when compared to the lead limb during STL (p ≤ .019). Greater trail limb knee flexion angles occurred during STL versus VJL at all four events (p ≤ .018), while greater plantarflexion angles occurred at all four events during VJL versus STL (p ≤ .034). During STL, greater trail limb plantarflexion angles were detected at each event versus the lead limb (p < .001). Lesser hip, lead and trail limb knee displacement occurred during STL versus VJL (p < .05). Greater vGRF impulse was detected during the loading phase of VJL (<.001), while greater vGRF impulse occurred during the attenuation phase of STL (p = .025). These tasks are characterized by distinct pre-contact kinematic strategies and post-contact kinetics. The task utilized in practice should reflect the requirements of the population of interest.


Aging asexual lineages and the evolutionary maintenance of sex.
Finite populations of asexual and highly selfing species suffer from a reduced efficacy of selection. Such populations are thought to decline in fitness over time due to accumulating slightly deleterious mutations or failing to adapt to changing conditions. These within-population processes that lead nonrecombining species to extinction may help maintain sex and outcrossing through species level selection. Although inefficient selection is proposed to elevate extinction rates over time, previous models of species selection for sex assumed constant diversification rates. For sex to persist, classic models require that asexual species diversify at rates lower than sexual species; the validity of this requirement is questionable, both conceptually and empirically. We extend past models by allowing asexual lineages to decline in diversification rates as they age, that is nonrecombining lineages "senesce" in diversification rates. At equilibrium, senescing diversification rates maintain sex even when asexual lineages, at young ages, diversify faster than their sexual progenitors. In such cases, the age distribution of asexual lineages contains a peak at intermediate values rather than showing the exponential decline predicted by the classic model. Coexistence requires only that the average rate of diversification in asexuals be lower than that of sexuals.


Target-specific cytotoxic activity of recombinant immunotoxin scFv(MUC1)-ETA on breast carcinoma cells and primary breast tumors.
MUC1 is a mucin family protein, overexpressed in more than 90% of breast cancers in an underglycosylated form, exposing the core peptides of the extracellular domain that act as a potential target for antibody-mediated therapy. We have developed an anti-MUC1 scFv antibody from a phage library of mice immunized with synthetic peptide MUC1-variable number of tandem repeats. MUC1 binding phages were affinity selected through biopanning using a biotin-streptavidin pull-down method. The selected phage clones showed target-specific binding to MUC1-expressing cells. Fusion of truncated Pseudomonas aeruginosa exotoxin A (ETA) to a high binder, phage-derived scFv clone and bacterial expression and purification of recombinant scFv(MUC1)-ETA immunotoxin were done with good yield and purity. In vitro target-specific cytotoxic activity and target-specific binding of immunotoxin were shown on MUC1-expressing cells and primary breast tumor samples. A truncated ETA fusion protein expressed from the same vector but lacking scFv did not show cytotoxic effects, confirming target specificity. Our results suggest that the scFv(MUC1)-ETA immunotoxin has therapeutic potential and deserves further development and characterization for MUC1-specific breast cancers treatment.


Is it still worth renewing nucleoside anticancer drugs nowadays?
Nucleoside has situated the convergence point in the discovery of novel drugs for decades, and a large number of nucleoside derivatives have been constructed for screening novel pharmacological properties at various experimental platforms. Notably, nearly 20 nucleosides are approved to be used in the clinic treatment of various cancers. Nevertheless, the blossom of synthetic nucleoside analogs in comparison with the scarcity of nucleoside anticancer drugs leads to a question: Is it still worth insisting on the screening of novel anticancer drugs from nucleoside derivatives? Hence, this review attempts to emphasize the importance of nucleoside analogs in the discovery of novel anticancer drugs. Firstly, we introduce the metabolic procedures of nucleoside anticancer drug (such as 5-fluorouracil) and summarize the designing of novel nucleoside anticancer candidates based on clinically used nucleoside anticancer drugs (such as gemcitabine). Furthermore, we collect anticancer properties of some recently synthesized nucleoside analogs, aiming at emphasizing the availability of nucleoside analogs in the discovery of anticancer drugs. Finally, a variety of synthetic strategies including the linkage of sugar moiety with nucleobase scaffold, modifications on the sugar moiety, and variations on the nucleobase structure are collected to exhibit the abundant protocols in the achievement of nucleoside analogs. Taken the above discussions collectively, nucleoside still advantages for the finding of novel anticancer drugs because of the clearly metabolic procedures, successfully clinic applications, and abundantly synthetic routines.


[Typing properties of bacteriophages isolated from Sh. sonnei strains].
On the basis of studying the lytic activity of eight bacteriophages the authors elaborated a scheme of bacteriophage typing of Sh. sonnei consisting of nine bacteriophage types with two subtypes. In typing 2348 strains isolated in various cities of the USSR bacteriophage pattern of Sh. sonnei was found to differ in different towns of the USSR. All the strains isolated repeatedly from the same patients were referred to the same bacteriophage type, this pointing to the stability of the type-determining properties of bacteriophages. Stability of bacteriophage types, a possibility of typing the strains of the I, II phases and of R-forms, simplicity and distinctness of the scheme of bacteriophage typing determined future prospects in using the type bacteriophages selected at the Tbilisi Institute of Vaccine and Sera for intraspecies differentiation of Sh. sonnei.


The Relationship between White Matter and Reading Acquisition, Refinement and Maintenance.
Reading requires efficient communication between brain regions that are situated all over the cortex. These brain areas are structurally connected by white-matter pathways that develop over the period of reading acquisition. Here, we conducted a cross-sectional study to assess the relationship between white-matter tracts and reading performance across the lifespan. Behavioral (reading performance) and neuroimaging (diffusion tensor imaging [DTI]) data were collected from participants aged 3-21 years as a part of the multi-site project called the pediatric imaging neurocognition genetics study. DTI measures (fractional anisotropy [FA], mean diffusivity, radial diffusivity and axial diffusivity) of bilateral dorsal tracts (arcuate fasciculus, superior longitudinal fasciculus, and posterior superior longitudinal fasciculus) and ventral tracts (uncinate fasciculus, inferior fronto-occipital fasciculus, and inferior longitudinal fasciculus) were extracted. Reading performance was calculated as the number of items correctly read (words for older children and letters in the case of young children). Correlational and regression analyses were conducted between the DTI measures and reading scores. During the early stages of reading acquisition (ages 3-6), the dorsal tracts were positively related to reading performance (as FA goes up, reading performance goes up). For ages 6-10, the dorsal tracts remain positively associated with reading performance and a relationship between ventral tracts and reading performance emerges. From the age of 10 onwards, the dorsal tracts no longer correlate with reading performance, and a brain-behavior relationship in the right ventral tracts begins to shift, whereby higher FA is associated with lower reading performance. In addition to the involvement of left hemispheric tracts, this study revealed the initial engagement of right hemispheric tracts during the early stages of reading acquisition. We discuss these different associations of white-matter tracts with reading during development in the context of the biological processes model of myelination and pruning.


The clinical effectiveness of a myofeedback-based teletreatment service in patients with non-specific neck and shoulder pain: a randomized controlled trial.
We investigated the effectiveness and efficiency of a four-week myofeedback-based teletreatment service in subjects with non-specific neck and shoulder pain. Subjects were recruited in Belgium, Germany and the Netherlands and randomly allocated to the intervention or conventional care. Subjects in the intervention group received four weeks of myofeedback training. Pain intensity and disability were evaluated by questionnaires at baseline, immediately after four weeks of treatment and at follow-up 3 months later. To investigate efficiency, the time-investment of both therapists and patients were assessed. Seventy-one subjects were included in the study (36 in the intervention group and 35 in the conventional care group). Myofeedback-based teletreatment was at least as effective clinically as conventional care. Pain intensity and disability decreased after 4 weeks of treatment in both groups and part of the effect remained at 3 months' follow-up. The teletreatment also increased efficiency for therapists by almost 20% and patients experienced the benefits of less travel time and travel costs by remote consultation. Myofeedback-based teletreatment has the potential to ensure more efficient treatment for patients with non-specific neck and shoulder pain.


Identification and validation of eight lysosomes-related genes signatures and correlation with immune cell infiltration in lung adenocarcinoma.
Lung cancer is the leading cause of cancer-related death. Lysosomes are key degradative compartments that maintain protein homeostasis. In current study, we aimed to construct a lysosomes-related genes signature to predict the overall survival (OS) of patients with Lung Adenocarcinoma (LUAD). Differentially expressed lysosomes-related genes (DELYs) were analyzed using The Cancer Genome Atlas (TCGA-LUAD cohort) database. The prognostic risk signature was identified by Least Absolute Shrinkage and Selection Operator (LASSO)-penalized Cox proportional hazards regression and multivariate Cox analysis. The predictive performance of the signature was assessed by Kaplan-Meier curves and Time-dependent receiver operating characteristic (ROC) curves. Gene set variant analysis (GSVA) was performed to explore the potential molecular biological function and signaling pathways. ESTIMATE and single sample gene set enrichment analysis (ssGSEA) were applied to estimate the difference of tumor microenvironment (TME) between the different risk subtypes. An eight prognostic genes (ACAP3, ATP8B3, BTK, CAV2, CDK5R1, GRIA1, PCSK9, and PLA2G3) signature was identified and divided patients into high-risk and low-risk groups. The prognostic signature was an independent prognostic factor for OS (HR > 1, p < 0.001). The molecular function analysis suggested that the signature was significantly correlated with cancer-associated pathways, including angiogenesis, epithelial mesenchymal transition, mTOR signaling, myc-targets. The low-risk patients had higher immune cell infiltration levels than high-risk group. We also evaluated the response to chemotherapeutic, targeted therapy and immunotherapy in high- and low-risk patients with LUAD. Furthermore, we validated the expression of the eight gene expression in LUAD tissues and cell lines by qRT-PCR. LYSscore signature provide a new modality for the accurate diagnosis and targeted treatment of LUAD and will help expand researchers' understanding of new prognostic models.


Burn resuscitation.
Fluid resuscitation following burn injury must support organ perfusion with the least amount of fluid necessary and the least physiological cost. Under resuscitation may lead to organ failure and death. With adoption of weight and injury size-based formulas for resuscitation, multiple organ dysfunction and inadequate resuscitation have become uncommon. Instead, administration of fluid volumes well in excess of historic guidelines has been reported. A number of strategies including greater use of colloids and vasoactive drugs are now under investigation to optimize preservation of end organ function while avoiding complications which can include respiratory failure and compartment syndromes. Adjuncts to resuscitation, such as antioxidants, are also being investigated along with parameters beyond urine output and vital signs to identify endpoints of therapy. Here we briefly review the state-of-the-art and provide a sample of protocols now under investigation in North American burn centers.


Electronic cigarettes: do they have a role in smoking cessation?
Electronic cigarettes have gained popularity among patients as a smoking cessation aid despite not being approved or supported for this purpose by the United States Food and Drug Administration due to concerns with poor manufacturing practices and the presence of known carcinogens in the limited products that they tested. A few studies have evaluated the effects of electronic cigarettes on plasma nicotine levels and heart rate but found negligible effects. Safety data are mainly limited to surveys in which patients report only minor side effects, such as mouth and throat irritation, headache, vertigo, and nausea. The efficacy of electronic cigarettes has been evaluated in studies in which patients report great success with being able to cut back or stop tobacco cigarette consumption. However, many of these studies introduce bias due to recruiting on e-cigarette Web sites and having tobacco cigarette use self-reported by the participant rather than objectively tested. A few studies have formally evaluated nicotine craving when using electronic cigarettes with mixed results. Although patients support the use of electronic cigarettes in smoking cessation, more formal studies on safety and efficacy should be completed in order to determine whether these products have a role in smoking cessation.


Deception by flexible alarm mimicry in an African bird.
Deception is common in nature, but victims of deception discriminate against and ultimately ignore deceptive signals when they are produced too frequently. Flexible variation of signals could allow evasion of such constraints. Fork-tailed drongos (Dicrurus adsimilis) use false alarm calls to scare other species away from food that they then steal. We show that drongos mimic the alarms of targeted species. Further, target species reduce their response to false alarm calls when they are repeated. However, the fear response is maintained when the call is varied. Drongos exploit this propensity by changing their alarm-call type when making repeated theft attempts on a particular species. Our results show that drongos can evade the frequency-dependent constraints that typically limit deception payoffs through flexible variation of their alarm calls.


The "Senobi" breathing exercise ameliorates depression in obese women through up-regulation of sympathetic nerve activity and hormone secretion.
Obese individuals have an increased risk of developing depression. This study aimed to determine whether the "Senobi" breathing exercise (SBE), a stretching-breathing exercise that we have established, could relieve depression, especially in obese women. Forty premenopausal women, aged 40 to 49 years, participated in the present study. Twenty were healthy, and the other 20 were obese (body mass index > 25 and body fat > 30%) and in a depressive state (OWD). Sympathetic nerve activity determined by analyzing heart rate variability, and the hormone levels in the urine were investigated before and 30 min after one minute of SBE. The relative proportion of sympathetic nerve activity among healthy women in the daytime was 79.2 ± 2.3%, whereas that in OWD group was 30.4 ± 1.9%. After one minute of SBE, significant up-regulation of sympathetic nerve activity and increased concentrations of catecholamines, estradiol, and growth hormone (all P values < 0.001) were observed in OWD group. After 30 days of SBE, the sympathetic nerve activity and hormone levels had recovered in OWD group, and the depressive state, as evaluated by the Hamilton Depression Scale, had ameliorated. The "Senobi" breathing exercise was found to be effective for amelioration of depression in obese women possibly through up-regulation of sympathetic nerve activity and hormone secretion.


A phototherapeutic fluorescent β-cyclodextrin branched polymer delivering nitric oxide.
We report herein on a novel water-soluble β-cyclodextrin-branched polymer covalently integrating a fluorescein moiety and a nitric oxide (NO) photodonor within its macromolecular skeleton. Photoexcitation with visible light induces the parallel activation of the two chromophores, which results in the green fluorescence emission suitable for imaging accompanied by NO release for therapy. In fact, this polymer internalizes in squamous carcinoma cancer cells in vitro, visualized by fluorescence microscopy, and induces cell mortality as result of the NO photo-decaging. The non-covalent drug delivery capability of this new material is also demonstrated using a hydrophobic photosensitizer for photodynamic therapy as a probe.


Synergy of Bacteroides fragilis and Escherichia coli in the induction of KC gene expression in mouse peritoneal tissues.
B. fragilis induces cytokine expression, which can serve as a host signal leading to inflammatory reaction and abscess formation in the peritoneal cavity. We assessed the hypothesis that enteric bacteria may alter the B. fragilis-induced expression of KC genes in mouse peritoneal tissues (MPT). After C57BL/6 mice were inoculated with abscess-forming mixture containing B. fragilis in the presence or absence of E. coli, RNA was extracted from MPT. Expression of KC mRNA was quantified using reverse-transcription polymerase chain reaction and standard RNA. KC and TNFalpha proteins were measured by enzyme-linked immunosorbent assay. KC mRNA in MPT was upregulated following inoculation of B. fragilis and this was paralleled by increased KC protein secretion. When the mice were co-infected with E. coli and B. fragilis intraperitoneally, there was a synergistic increase in the expression of KC of MPT. Co-infection with L. acidophilus and B. fragilis downregulated KC mRNA expression, but co-infection with E. faecalis and B. fragilis synergistically increased KC expression in the infected MPT. Inhibition of TNFalpha production could downregulate KC expression in mixed infected MPT. These results suggest that enteric bacteria may significantly affect the KC signal produced by the host peritoneal cavity in response to B. fragilis infection.


Comparison of the effects of clinical observation and protocol-based weaning on antioxidant stress factors.
We aimed to compare the effects of observation of the physician (POB) or by adhering to the protocol-based (PB) weaning methods on total antioxidant capacity (TAC) and total oxidative stress (TOS) levels and weaning success levels. Our study was conducted on patients admitted from the emergency department between January 2015 and January 2018 in the intensive care unit of our hospital. During the spontaneous breathing trial (SBT), when one of the criteria specified in developed, SBT was terminated and the previous mechanical ventilator parameters were returned. The patient was planned to be taken to SBT again the next morning. If the SBT was successful, extubation was decided. The extubation decision based on physician observation was made according to the patient's state of consciousness and adequate chest expansion during the daily visit. The decrease in TAC average value before and after extubation was found to be significant in the POB group patients (p=0.001). The decrease in the average TAC value of the PB group patients before and after extubation was found to be significant (p=0.03). In our study, TAC values were found to be higher in the PB group than in the POB group, and in addition, the reintubation rate was found to be lower. We think that the management of weaning as a PB may contribute to maintaining the balance between TAC and TOS and reduce the rate of reintubation.


Evaluating the impact of eligibility for free care on the use of general practitioner (GP) services: a difference-in-difference matching approach.
In Ireland, approximately 30% of the population ('medical card patients') are entitled to free general practitioner (GP) care while the remaining 70% ('private patients') must pay the full cost of each visit. Previous research has analysed the effect of this system on GP visiting patterns using regression methods, but to date, no attempt has been made to apply techniques from the treatment evaluation literature to this issue. Treatment evaluation techniques are commonly employed when observations are not randomly assigned to treatment and control groups; this is certainly the case here, as the primary criterion for medical card eligibility is an income below a specified income threshold (and individuals may also be granted medical cards for other reasons such as chronic ill-health). In this paper, previous Irish research, which has analysed the effect of medical card eligibility on GP visiting using regression methods, is extended to consider the use of difference-in-difference matching methods, which control for non-random selection into treatment and control groups, as well as differences in time-invariant unobserved characteristics between individuals in both groups. The results are largely consistent with earlier results using pooled cross-sectional and panel data, and confirm that medical card eligibility exerts a significant effect on GP visiting, even after controlling for observed and unobserved differences in characteristics between medical card and private patients.


Less educated and older patients have reduced access to biologic DMARDs even in a country with highly developed social welfare (Norway): results from Norwegian cohort study NOR-DMARD.
To explore whether age, gender or education influence the time until initiation of the first bDMARD in patients with RA. Data from the Norwegian Register of DMARDs collected between 2000 and 2012 were used. Only DMARD-naïve patients with RA starting their first conventional synthetic DMARD were included in the analyses. The start of the first bDMARD was the main outcome of interest. Cox regression analyses were used to explore the impact of education, age and gender on the start of a first bDMARD, adjusting for confounders, either at baseline or varying over time (time-varying model). Of 1946 eligible patients [mean (s.d.) age: 55 (14) years, 68% females], 368 (19%) received a bDMARD during follow-up (mean 2.6 years). In the baseline prediction model, older age [Hazard Ratio (HR) 0.97, 95% CI: 0.96, 0.98], lower education [HR = 0.76 and 0.68 for low and intermediate education levels vs college/university education, respectively (P = 0.01)] and female gender [only in the period 2000-03, HR = 0.61 (95% CI: 0.41, 0.91)] were associated with a lower hazard ratio to start a bDMARD. The time-varying model provided overall consistent results, but the effect of education was only relevant for older patients (>57 years) and became more pronounced by the end of the decade. Less educated and older patients have disadvantages with regard to access to costly treatments, even in a country with highly developed welfare like Norway. Females had lower access in the beginning of the 2000s, but access had improved by the end of the decade.


The role of front-line anthracycline-containing chemotherapy regimens in peripheral T-cell lymphomas.
Peripheral T-cell lymphomas (PTCLs) are a heterogenous group of aggressive non-Hodgkin's lymphomas that are incurable in the majority of patients with current therapies. Outcomes associated with anthracycline-based therapies are suboptimal, but remain the standard of care for most patients, even though the benefits of this approach remain uncertain. This study retrospectively examined outcomes in a cohort of North American PTCL patients treated with both anthracycline- and nonanthracycline-containing regimens. The incorporation of anthracycline-containing regimens was associated with improved progression-free survival (PFS) and overall survival (OS). Patients treated with nonanthracycline-containing regimens were more likely to have high-risk features and were less likely to undergo high-dose therapy and stem cell transplantation. However, anthracycline use remained an independent predictor of improved PFS and OS when adjusting for these confounding variables. Anthracycline-based regimens and consolidation with high-dose therapy and autologous stem cell transplantation in appropriately selected patients remains a viable option for patients unable to participate in a clinical trial. Long-term disease-free survival is not optimal, highlighting the need for an improved understanding of disease pathogenesis, and the development of novel therapeutic strategies.


Preterm, early term, and post-term infants from Riyadh mother and baby multicenter cohort study: The cohort profile.
Birth before 37 or beyond 42 gestational weeks is associated with adverse neonatal and maternal outcomes. Studies investigating determinants and outcomes of these deliveries are scarce. The objective of this study was to determine the neonatal birth profile in relation to the gestational age at delivery and to evaluate its influence on the immediate maternal and neonatal outcomes. This is a multicenter cohort study of 13,403 women conducted in three hospitals in Riyadh. Collected data included sociodemographic characteristics, obstetric history, and physical and laboratory measurements. Regression models were developed to estimate the adjusted odds ratio (OR) and confidence intervals (CI) to determine factors associated with preterm, early term, and post-term births and to evaluate common maternal and neonatal risks imposed by deliveries outside the full term. The incidence of preterm, early term, and post-term delivery was 8.4%, 29.8%, and 1.4%, respectively. Hypertensive events during pregnancy consistently increased the risk of all grades of preterm births, from more than 3-fold for late preterm (OR = 3.40, 95% CI = 2.21-5.23) to nearly 7-fold for extremely early preterm (OR = 7.11, 95% CI = 2.24-22.60). Early term was more likely to occur in older mothers (OR = 1.30, 95% CI = 1.13-1.49), grand multiparous (OR = 1.21, 95% CI = 1.06-1.38), pregestational diabetes (OR = 1.91, 95% CI = 1.49-2.44), and gestational diabetes women (OR = 1.18, 95% CI = 1.05-1.33). The risk of post-term birth was higher in primiparous. In preterm births, the adverse outcome of neonates having an APGAR score of <7 at 5 min and admission to neonatal intensive care units increased progressively as the gestational age decreased. Post-term births are 2-fold more likely to need induction of labor; meanwhile, preterm births were more likely to deliver by cesarean section. This large cohort study was the first in Saudi Arabia to assess the delivery profile across a continuum of gestational age and the associated maternal and neonatal adverse outcomes of deliveries outside the full-term period. The study showed that the prevalence of preterm and post-term birth in Saudi Arabia is similar to the prevalence in other high-income countries. The immediate adverse pregnancy outcomes inversely increased with the decrease in gestational age at delivery. In addition, maternal age, hypertension, diabetes, and parity influenced the gestational age at delivery.


Comparison of the specific incorporation of intracrystalline proteins into urinary calcium oxalate monohydrate and dihydrate crystals.
The aim of this study was to compare the comprehensive intracrystalline protein profiles of calcium oxalate monohydrate (COM) and dihydrate (COD) crystals precipitated from the same human urine samples. Three separate batches of COM and COD crystals were precipitated from pooled healthy human urine by the addition of sodium oxalate at calcium concentrations of 2 and 8 mM, respectively. Proteins in whole extracts of demineralised COM and COD crystals, as well as in spots excised from 2D-PAGE gels of the extracts, were identified using liquid chromatography and tandem mass spectrometry (LC-MS/MS). The number and type of individual proteins differed between COM and COD: 14 substantive proteins were found inside COM crystal extracts and 34 inside COD, with 9 proteins occurring in both crystal types. Numerous keratins were detected. However, in line with consensus in the proteomics literature, as well as a lack of published evidence linking them to urolithiasis, they were excluded as contaminants, leaving very few consistently detected proteins. On the basis of their known association with stone disease or identification in multiple runs, the principal proteins in COM crystal extracts were prothrombin fragment 1, protein S100A9, and IGkappaV1-5, while those in extracts of COD crystals included osteopontin, IGkappaV1-5, protein S100A9, annexin A1, HMW kininogen-1, and inter-alpha-inhibitor (IalphaI). In general, proteins incorporated into both hydromorphs were acidic (pI<6), smaller than 55 kDa, and calcium binders. We concluded that the incorporation of proteins into urinary COM and COD crystals is selective and that only a few of the urinary proteins associated with the two hydromorphs are likely to play any significant role in stone pathogenesis.


Anti-Mycobacterium paratuberculosis (MAP) therapy for Crohn's disease: an overview and update.
The role of Mycobacterium avium subspecies paratuberculosis (MAP) in the pathogenesis of Crohn's disease (CD) has been strongly debated for many years. MAP is the known aetiological agent of Johne's disease, a chronic enteritis affecting livestock. At present, due to the paucity of high-quality data, anti-MAP therapy (AMT) is not featured in international guidelines as a treatment for CD. Although the much-quoted randomised trial of AMT did not show sustained benefits over placebo, questions have been raised regarding trial design, antibiotic dosing and the formulation used. There are several lines of evidence supporting the CD and MAP association with uncontrolled and controlled trials demonstrating effectiveness, including a retrospective review of cases treated at our own institution. Here, we provide an overview of the evidence supporting and refuting AMT in CD before focussing on updates of the current research in the field, including the ongoing trials with the novel RHB-104 formulation and the MAP vaccine trial. While controversial, gastroenterologists are often asked about long-term combination antibiotic therapy for CD. There has been broadcast and social media coverage surrounding this, particularly with regard to current trials. Although patients should not be deterred from treatments of proven effectiveness, this review aims to help with commonly asked questions and highlights our own approach for the use of anti-MAP in specific circumstances.


The Smith-Lemli-Opitz syndrome.
The Smith-Lemli-Opitz syndrome (SLOS) is one of the archetypical multiple congenital malformation syndromes. The recent discovery of the biochemical cause of SLOS and the subsequent redefinition of SLOS as an inborn error of cholesterol metabolism have led to important new treatment possibilities for affected patients. Moreover, the recent recognition of the important role of cholesterol in vertebrate embryogenesis, especially with regard to the hedgehog embryonic signalling pathway and its effects on the expression of homeobox genes, has provided an explanation for the abnormal morphogenesis in the syndrome. The well known role of cholesterol in the formation of steroid hormones has also provided a possible explanation for the abnormal behavioural characteristics of SLOS.


Type II cAMP-dependent protein kinase is associated with the rabbit kidney brush border membranes.
Photolabelling with 32P-8-azido-cAMP identified a major cAMP-binding protein (54 kDa) in isolated rabbit renal apical membranes, whose labelling was competitively inhibited by cAMP. Membrane associated cAMP-binding polypeptides were extensively purified by affinity chromatography on cAMP-Sepharose. The 54 kDa polypeptide represented 70-80% of the total protein eluted with cAMP. This protein was rapidly phosphorylated by the catalytic subunit of cAMP-dependent protein kinase, with a shift in its apparent mobility on SDS-PAGE to Mr 56/58,000. The phosphopeptide maps of autophosphorylated rat skeletal muscle RII and rabbit kidney 56/58 kDa proteins were essentially identical. Western immuno-blot analysis, using antibodies generated against purified rat RI and RII, indicated preferential cross-reactivity of rabbit kidney 54 kDa protein with anti-RII antibodies. The data demonstrates the specific association of the regulatory subunit of type II cAMP dependent protein kinase with rabbit renal brush border membranes.


Child drawings and salivary cortisol in children undergoing preoperative procedures associated with day surgery.
Perioperative procedures in children can impair their emotional status negatively with stress and/or anxiety. Cortisol concentrations and drawings could be helpful in gaining information about a child's levels of stress and/or anxiety when attending the hospital for surgery. The purpose of this study was to determine the degree of anxiety and stress as well as to explore the association between objective measures of stress (cortisol concentration in saliva) and subjective assessment of hospital anxiety (children's drawings) as interpreted by the Swedish version of the Child Drawing: Hospital manual. A total of 93 children scheduled for day surgery were included. Salivary cortisol was sampled preoperatively on the day of surgery at which time the children were also requested to make a drawing of a person at the hospital. Results showed no association between salivary cortisol concentration and the CD:H score. The drawings and salivary cortisol concentration preoperatively on the day of surgery reflect different components of the conditions of fear, anxiety, or stress emerging in the situation.


The clinical value of the Duke Anesthesia Resistance Scale in predicting postoperative delirium after hip fracture surgery: a retrospective study.
This study aims to investigate the clinical value of the Duke Anesthesia Resistance Scale (DARS) in predicting postoperative delirium (POD) after hip fracture surgery. A retrospective study was conducted. Clinical data were collected from the patients who had hip fracture and underwent elective total hip arthroplasty in Shaanxi Provincial People's Hospital, Third Affiliated Hospital of Xi'an Jiaotong University between January 2022 and June 2023. The Consciousness Fuzzy Assessment Scale was used to evaluate the occurrence of POD on postoperative day 3 (POD 3). The enrolled patients were divided into the POD group (n = 26) and the non-POD group (n = 125). Baseline characteristics, surgical data, postoperative information, and laboratory test results were collected. DARS scores were calculated using the minimum alveolar concentration, end-tidal concentration average (ETAC), and bispectral index (BIS). Multivariate logistic regression analysis was conducted to recognize the independent risk factors for POD after hip fracture surgery. Receiver operating characteristic (ROC) curve was plotted to evaluate the value of DARS in POD prediction. The average age of POD group was significantly higher, comparing to non-POD group (P < 0.05). DARS scores were statistically lower in the POD group compared to non-POD group (P < 0.05). Multivariate logistic regression analysis found that age and DARS scores were factors impacting post-operative delirium occurrence after hip fracture surgery (P < 0.05). ROC showed that the area under the curve for DARS in predicting POD after hip fracture surgery was 0.929 (95% CI [0.861-0.997]). The optimal cutoff value was 30. The sensitivity was 95.45%, while the specificity was 84.09%. DARS score demonstrates good predictive value in hip fracture patients and is feasible in clinical practice, making it suitable for clinical application and promotion.


Interpersonal Discrimination, Neighborhood Inequities, and Children's Body Mass Index: A Descriptive, Cross-Sectional Analysis.
Psychosocial stressors have been implicated in childhood obesity, but the role of racism-related stressors is less clear. This study explored associations between neighborhood inequities, discrimination/harassment, and child body mass index (BMI). Parents of children aged 5-9 years from diverse racial/ethnic backgrounds (n = 1307), completed surveys of their child's exposure to discrimination/harassment. Census tract data derived from addresses were used to construct an index of concentration at the extremes, a measure of neighborhood social polarization. Child's height and weight were obtained from medical records. Multiple regression and hierarchical models examined child's BMI and racism at the individual and census tract levels. Children residing in the most Black-homogenous census tracts had 8.2 percentage units higher BMI percentile (95% confidence interval, 1.5-14.9) compared with white-homogenous tracts (P = .03). Household income and home values were lower, poverty rates higher, and single parent households more common among Black-homogeneous census tracts. Almost 30% of children experienced discrimination/harassment in the past year, which was associated with a 5.28-unit higher BMI percentile (95% confidence interval, 1.72-8.84; P = .004). Discrimination and racial/economic segregation were correlated with higher child BMI. Longitudinal studies are needed to understand whether these factors may be related to weight gain trajectories and future health.


Functional consequences of interleukin 2 receptor expression on resting human lymphocytes. Identification of a novel natural killer cell subset with high affinity receptors.
In this study, we have used radiolabeled IL-2 binding assays, Northern blot analysis, immunofluorescent flow cytometry and cell sorting, as well as proliferation and cytotoxicity assays to perform an extensive phenotypic and functional characterization of the IL-2 receptor in normal resting human peripheral blood lymphocytes. Our results indicate that almost all T cells (greater than 98%) express neither the high affinity IL-2 receptor nor the functional intermediate affinity p75 chain of the IL-2 receptor without prior activation. In contrast, most NK cells constitutively express the isolated intermediate affinity p75 IL-2 receptor. In addition, a subpopulation of NK cells, distinguished by high density expression of the NKH1 antigen, constitutively express the high affinity IL-2 receptor, in addition to an excess of the isolated intermediate affinity p75 IL-2 receptor. These NKH1bright+ cells exhibit a brisk proliferative response to IL-2, similar to that seen with antigen-activated T cells, yet do so in the absence of any known antigenic stimuli. No other resting peripheral blood lymphocyte population, including CD4+, CD8+, and CD20 cells, exhibits this property. The intermediate affinity p75 IL-2 receptor, as it exists in its isolated form on resting NK cells, does not transduce a growth signal equivalent to that seen in NK cells expressing the high affinity IL-2 receptor, despite doses of IL-2 that are known to fully saturate the isolated p75 chain. This strongly suggests that additional structural or functional components are involved in generating the proliferative response following the binding of IL-2 to the high affinity heterodimeric form of the IL-2 receptor. The constitutive expression of this functional high affinity IL-2 receptor on a small population of resting NK cells provides further evidence in support of a role for these cells in the host's early defense against viral infection or malignant transformation, before the more delayed but specific T cell response.


Evaluation and modification of the overall hemostasis potential assay for use with canine plasma.
To optimize the overall hemostasis potential (OHP) assay for use with canine platelet-poor plasma and determine reference intervals in healthy dogs. 40 healthy dogs. Blood was collected from the dogs into citrated tubes, and platlet-poor plasma was obtained. The OHP assay and standard coagulation assays (prothrombin time, activated partial thromboplastin time, and fibrinogen concentration) were performed for each sample. The OHP assay outputs were tested for correlations with results of the standard coagulation assays, age, and sex. Modifications to the published methodology for the OHP assay were required for use with canine plasma, with less coagulation activator (thrombin) and more fibrinolysis activator (tissue plasminogen activator) than used with human plasma. Male dogs had a higher OHP than did females. High fibrinogen concentrations were associated with increases in maximum optical density, OHP, and overall coagulation potential, and reduced prothrombin time was associated with increases in maximum optical density, overall coagulation potential, OHP, and maximum slope. Results supported the use of the OHP assay as an accessible, cost-effective global coagulation assay. Further research is required to determine its clinical application as an alternative to thromboelastography or thrombin generation assays.


Risk factors for challenging behaviour and psychopathology in adults with Down syndrome.
This study explored the role of demographic, psychosocial, stress and life event-related risk factors for psychopathology and challenging behaviour in a clinical sample of adults with Down syndrome. A convenience sample of adults with Down syndrome seen through a specialised clinic was assessed through interviews and questionnaires specifically designed for use in people with intellectual disabilities. Recent negative life events and stressors were significantly correlated with mental and behavioural health, and significantly predicted irritability, lethargy and depressed mood. Social avoidance was predicted by stress related to anticipation and social-environmental stressors, and negatively predicted by verbal ability. Ritual-related stress predicted obsessive-compulsive behaviour. Participants who did not have a job or vocational placement were significantly more depressed than participants who did. Adults with Down syndrome should be provided increased supports for coping with negative life events and stressors. Interventions should also emphasise community engagement, such as employment, and access to psychosocial supports that teach coping and self-regulation skills when faced with stressors and negative life events.


[Clear lens exchange should be performed with caution].
Clear lens exchange (CLE) boasts a venerable lineage, its prominence in the field of ophthalmology having incrementally solidified over recent decades, concomitant with the evolution of surgical techniques and the innovation of intraocular lens materials. CLE has assumed a pivotal role in the therapeutic armamentarium for conditions such as glaucoma, high myopia, and presbyopia. Nevertheless, it remains that the sacrifice of accommodation, potentialities of intraoperative and postoperative complications, and less than optimal results in terms of spectacle independence resulting from CLE are not infrequent occurrences. The indiscriminate pursuit of this surgical intervention is inadvisable. It is only by means of a sagacious appreciation of the merits and demerits of CLE, the judicious circumscription of its indicatory ambit, and the bespoke selection of surgical modalities catering to the individual patient, that the superlative outcomes in the realms of surgical efficacy, safety, and patient contentment may be attained.


Iron Carbonate Beneficiation Through Reductive Calcination - Parameter Optimization to Maximize Methane Formation.
Direct iron carbonate reduction through reductive calcination in a hydrogen atmosphere is a high-potential candidate for environmentally benign pig iron production. In addition to the direct formation of elemental iron in one reaction step, carbon dioxide is only partially released from the carbonate. Instead, carbon monoxide, methane, and higher hydrocarbons form as gaseous reaction products. The experimental study described here is based on Mg-Mn substituted iron carbonate ore. First, the chemical thermodynamics of the reductive calcination of iron, magnesium, and manganese carbonate are discussed. The influence of temperature and pressure on equilibrium conversion is reviewed together with the accessible products. Results for the reductive calcination of mineral iron carbonate in a tubular reactor setup are presented. The methane yield was optimized via statistically planned design of experiments. The gauge pressure and temperature showed a statistically significant effect on the total iron carbonate conversion, as well as on carbon monoxide, and methane yield.


Relationship of hepatic functional imaging to irinotecan pharmacokinetics and genetic parameters of drug elimination.
The marked variability of irinotecan (Ir) clearance warrants individualized dosing based on hepatic drug handling. The aims of this trial were to identify parameters from functional hepatic nuclear imaging (HNI) that correlate with (1) Ir pharmacology, and (2) single-nucleotide polymorphisms (SNPs) for the ABCB1 (P-glycoprotein) and UGT-1A1 genes, known to influence Ir handling. Patients underwent genotyping for ABCB1 SNPs and UTUGT-1A1*28 carriage and HNI with 99mTc-DIDA (acetanilidoiminodiacetic acid)/99mTc-DISIDA (disofenin) and MIBI (99mTc-sestamibi) scans, probes for biliary transport proteins ABCC1 and -2, and ABCB1 function. HNI data were analyzed by noncompartmental and deconvolutional analysis to provide hepatic extraction and biliary excretion parameters. Patients received Ir, fluorouracil, and folinic acid using a weekly x2, every-3-weeks schedule. Plasma was taken for Ir and SN-38 analysis on day 1, cycle 1. Of the 21 patients accrued, Ir pharmacokinetics data were obtained from 16 patients. 99mTc-DIDA/DISIDA percent retention at 1 hour (1-hour RET) correlated to baseline serum bilirubin (P = .008). Both 99mTc-DIDA/DISIDA and MIBI 1-hour RET correlated with SN-38 area under the curve (AUC; P < .01). On multiple regression analysis, SN-38 AUC = -215 + 18.68 x bilirubin + 4.27 x MIBI 1-hour RET (P = .009, R2 = 44.2%). HNI parameters did not correlate with Ir toxicity or UGT1A1*28 carriage. MIBI excretion was prolonged in patients with the ABCB1 exon 26 TT variant allele relative to wild-type (P = .015). Functional imaging of hepatic uptake and excretory pathways may have potential to predict Ir pharmacokinetics. Evaluation of a larger cohort as well as polymorphisms in other biliary transporters and UGT1A1 alleles is warranted.


Investigation of two different size microplastic degradation ability of thermophilic bacteria using polyethylene polymers.
There are several studies stating that many types of microplastics cannot be retained completely by conventional wastewater treatment systems. Therefore, it is necessary to prevent the discharge of these microplastics to the ecological system. The objective of this study was to investigate the biodegradation ability of two different size of PE (50 and 150 µm) by using two Gram-positive, spore-forming, rod-shaped, and motile thermophilic bacteria, called strain Gecek4 and strain ST5, which can hydrolyse starch, were isolated from the soil's samples of Gecek and Ömer hot-springs in Afyonkarahisar, Turkey, respectively. Phenotypic features and 16S rRNA analyzing of strains also studied. According to these results, Gecek4s and ST5 were identified as Anoxybacillus flavithermus Gecek4s and Bacillus firmus ST5, respectively. Results showed that A. flavithermus Gecek4s could colonise the polymer surface and cause surface damage whereas B. firmus ST5 could not degrade bigger-sized particles efficiently. In addition, morphological changes on microplastic surface were investigated by scanning electron microscopy (SEM) where dimensional changes, irregularities, crack, and/or holes were detected. This finding suggests that there is a high potential to develop an effective integrated method for plastic bags degradation by extracellular enzymes from bacteria.


[Placentary chorangio-leiomyoma].
Chorangiomas are the most common non-trophoblastic tumors, generally appearing as incidental findings at the moment of delivery. The presence of leiomyomas inside placental parenchyma has been also described sporadically. In these cases, leiomyomas were primary maternal uterine neoplasms incorporated into the placenta during pregnancy. This case report presents a very unusual case of mesenchymal lesion, located in the fetal surface of the placenta, characterized by combined histopathologic and immunohistochemical features of chorangioma and leiomyoma. A single case reported with these characteristics was found in the international scientific literature, named as "chorangioleiomyoma". To our knowledge, this is the first case reported in our country.


Regulation of inflammation, antioxidant production, and methyl-carbon metabolism during methionine supplementation in lipopolysaccharide-challenged neonatal bovine hepatocytes.
Supplementation of methionine (Met) may improve immunometabolic status, specifically during a period of inflammatory stress. The aim of the present study was to establish an inflammation model using primary neonatal bovine hepatocytes and to examine the effects of increasing concentrations of dl-Met and a maintained Met to lysine (Lys) ratio on hepatocyte inflammatory responses, antioxidant production, and Met metabolism during lipopolysaccharide (LPS) challenge. Hepatocytes isolated from 4 calves were maintained as monolayer cultures and exposed to 0, 10, or 40 µMdl-Met and 100 µM Lys (0Met100Lys, 10Met100Lys, or 40Met100Lys) or 10 µMdl-Met and 25 µM Lys (10Met25Lys). Cells were exposed to each treatment for 16 h and then challenged with either 0 or 100 ng/mL of LPS for 8 h. In the absence of LPS, glutathione (GSH) was not altered by 10Met100Lys or 10Met25Lys but was increased by 40Met100Lys. With LPS challenge, GSH concentration was decreased with 40Met100Lys and tended to be decreased with 10Met100Lys. Hepatocytes receiving 10Met100Lys treated with 100 ng/mL of LPS showed an inflammatory response with increased mRNA expression of tumor necrosis factor (TNFα), IL-6, IL-1β, and interferon gamma, which was accompanied by increased nuclear factor κB inhibitor and serum amyloid A3 mRNA. The treatment 40Met100Lys was effective for preventing the LPS-induced increase in expression of the above genes except TNFα. Similar preventative effects were observed for 10Met25Lys; however, it did not prevent the LPS-induced increase in TNFα or IL-6 mRNA. Lipopolysaccharide challenge decreased mRNA expression of key genes controlling the transmethylation and Met regeneration pathways, which was not prevented by Met supplementation. The data suggest that bovine hepatocyte cultures can be used as a biological model to study the inflammatory cascade via an LPS challenge. Supplementation of Met prevents the LPS-induced hepatocyte cytokine expression and is associated with elevated intracellular GSH concentration.


Reduced adipose tissue H2S in obesity.
Hydrogen sulfide (H2S) is an endogenously produced signaling molecule synthesized by cystathionine γ-lyase (CSE), cystathionine β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (3-MST). Given that H2S exerts significant effects on bioenergetics and metabolism, the goal of the current study was to determine the expression of H2S-producing enzymes in adipose tissues in models of obesity and metabolic disruption. Mice fed a western diet expressed lower mRNA levels of all three enzymes in epididymal fat (EWAT), while only CSE and 3-MST were reduced in brown adipose tissue (BAT). At the protein level 3-MST was reduced in all fat depots studied. Using db/db mice, a genetic model of obesity, we found that CSE, CBS and 3-MST mRNA were reduced in white fat, while only CSE was reduced in BAT. CBS and CSE protein levels were suppressed in all three fat depots. In a model of age-related weight gain, no reduction in the mRNA of any of the enzymes was noted. Smaller amounts of 3-MST protein were found in EWAT, while both CSE and 3-MST were reduced in BAT. Tissue levels of H2S were lower in WAT in HFD mice; both WAT and BAT contained lower H2S amounts in db/db animals. Taken together, our data suggest that obesity is associated with a decreased expression of H2S-synthesizing enzymes and reduced H2S levels in adipose tissues of mice. We propose that the reduction in H2S may contribute to the metabolic response associated with obesity. Further work is needed to determine whether restoring H2S levels may have a beneficial effect on obesity-associated metabolic alterations.


Extracorporeal Removal of Poisons and Toxins.
Extracorporeal therapies have been used to remove toxins from the body for over 50 years and have a greater role than ever before in the treatment of poisonings. Improvements in technology have resulted in increased efficacy of removing drugs and other toxins with hemodialysis, and newer extracorporeal therapy modalities have expanded the role of extracorporeal supportive care of poisoned patients. However, despite these changes, for at least the past three decades the most frequently dialyzed poisons remain salicylates, toxic alcohols, and lithium; in addition, the extracorporeal treatment of choice for therapeutic removal of nearly all poisonings remains intermittent hemodialysis. For the clinician, consideration of extracorporeal therapy in the treatment of a poisoning depends upon the characteristics of toxins amenable to extracorporeal removal (e.g., molecular mass, volume of distribution, protein binding), choice of extracorporeal treatment modality for a given poisoning, and when the benefit of the procedure justifies additive risk. Given the relative rarity of poisonings treated with extracorporeal therapies, the level of evidence for extracorporeal treatment of poisoning is not robust; however, extracorporeal treatment of a number of individual toxins have been systematically reviewed within the current decade by the Extracorporeal Treatment in Poisoning workgroup, which has published treatment recommendations with an improved evidence base. Some of these recommendations are discussed, as well as management of a small number of relevant poisonings where extracorporeal therapy use may be considered.


Mitochondrial DNA deletions are associated with non-B DNA conformations.
Mitochondrial DNA (mtDNA) deletions are a primary cause of mitochondrial disease and are believed to contribute to the aging process and to various neurodegenerative diseases. Despite strong observational and experimental evidence, the molecular basis of the deletion process remains obscure. In this study, we test the hypothesis that the primary cause of mtDNA vulnerability to breakage resides in the formation of non-B DNA conformations, namely hairpin, cruciform and cloverleaf-like elements. Using the largest database of human mtDNA deletions built thus far (753 different cases), we show that site-specific breakage hotspots exist in the mtDNA. Furthermore, we discover that the most frequent deletion breakpoints occur within or near predicted structures, a result that is supported by data from transgenic mice with mitochondrial disease. There is also a significant association between the folding energy of an mtDNA region and the number of breakpoints that it harbours. In particular, two clusters of hairpins (near the D-loop 3'-terminus and the L-strand origin of replication) are hotspots for mtDNA breakage. Consistent with our hypothesis, the highest number of 5'- and 3'-breakpoints per base is found in the highly structured tRNA genes. Overall, the data presented in this study suggest that non-B DNA conformations are a key element of the mtDNA deletion process.