Endovascular intervention for acute cervical carotid artery occlusion.
Acute cervical carotid artery occlusion presents with a severe neurological deficit and is associated with unfavorable outcomes. In this study, the authors report their experience with patients having had acute ischemic stroke due to cervical carotid occlusion, who underwent endovascular intervention. Sixteen acute cervical carotid occlusion patients (15 males and 1 female; mean age 67.7 years) were treated by endovascularly between January 2009 and November 2012. Clinical, procedural, and angiographic data were retrospectively evaluated. Successful intracranial recanalization was based on thrombolysis in cerebral infarction score of 2B-3. A favorable outcome was defined as a modified Rankin Scale score of 0-2 at 90 days. The average score of National Institutes of Health Stroke Scale before treatment was 15.9. Ten of 16 patients (63 %) were associated with intracranial tandem occlusion. Ten (63 %) cases were caused by atherosclerotic, 4 (25 %) by atrial fibrillation (AF), and 2 (13 %) by dissection. Thirteen of 16 (81 %) achieved successful cervical recanalization and 7 of 16 (44 %) patients obtained sufficient cervical and intracranial perfusion. As a result, 5 of 16 (31 %) patients demonstrated favorable outcomes. Five of seven patients (71 %) with successful cervical and intracranial recanalization presented favorable outcomes. In contrast, none of the patients without cervical or intracranial recanalization presented favorable outcomes. Three of 6 (50 %) patients initially without intracranial occlusion showed favorable outcomes, but only 2 of 10 (20 %) patients associated with intracranial occlusion had favorable outcomes. On the aspect of etiology, in atherosclerotic cases, 4 of 10 (40 %) showed favorable outcomes. However, all four AF cases deteriorated into poor outcomes. This study demonstrated the feasibility of endovascular intervention for acute cervical carotid artery occlusion. Although treatment for tandem occlusion and AF cases is an issue that should be resolved, intervention must be encouraged. Successful cervical and intracranial revascularization will be essential for favorable outcomes.


The effects of 4-aminopyridine and methylprednisolone on recovery of the facial nerve crush injury.
4-Aminopyridine (4-AP) is a potassium channel blocker that enhances nerve excitability. In this study, rat models that have facial nerve crush injury (FNCI) were grouped and treated with methylprednisolone (MP), 4-AP, and a combination of these two drugs. Electrophysiologic and histopathologic outcomes of these groups will be compared with a control group. Thirty healthy male Wistar rats (mean weight of 265 g) were used in this study. The rats were randomly divided into five groups with six subjects in each: Group 1 (sham group), Group 2 (control group), Group 3 (MP group), Group 4 (4-aminopyridine group), and Group 5 (4-AP + MP group). All groups except the sham group underwent crush injury to the right facial nerve. Electrophysiologic and histologic recovery was recorded three weeks postoperatively. The 4-AP group and the combined group had a more significant recovery at Nerve Excitability Thresholds (NET) at the end of three weeks. The methylprednisolone group and the control group had a minimal recovery of NET. Histologically, when compared with the control group, the combined group was the only group that had significant recovery at all three of axonal degeneration, axon diameter, and myelin thickness. In this experimental study, we demonstrated that a combination treatment of 4-AP and MP is more effective in the recovery of peripheric FNCI than in the no-treatment control group and in the 4-AP- or MP-alone groups. Moreover, our results suggested that 4-AP can be a potent alternative to MP in the treatment of the FNCI. N/A.


Arg124Cys mutation of the betaig-h3 bene in a Japanese family with lattice corneal dystrophy type I.
To characterize severe lattice corneal dystrophy, we analyzed the betaig-h3 gene, clinical features, histological findings, and genotype-phenotype correlation in an affected Japanese family. Deoxyribonucleic acid was extracted from leukocytes in 16 members (12 affected and 4 unaffected) of a Japanese family with lattice corneal dystrophy type I. Exon 4 of the betaig-h3 gene was amplified and analyzed using molecular biological methods. Clinical and pathological data were also collected. We found a heterozygous point mutation that causes the disease phenotype. It was a single base-pair transition leading to an amino acid substitution (CGC-->TGC, Arg124Cys). The phenotypic variation within families was not recognized. The affected members in the pedigree demonstrated severe visual disturbance in the third decade and required keratoplasty. Histopathological examination revealed amyloid deposits consisting of short and thin amyloid fibers and lattice corneal dystrophy type I. The heterozygous Arg124Cys mutation reported in Caucasian lattice corneal dystrophy caused severe lattice corneal dystrophy consisting of short and thin amyloid fibers in a Japanese family. Based on our study of many members of the family, we are able to construct the natural course of this disorder from its earliest clinical findings through its late manifestations.


Combined effects of zearalenone and deoxynivalenol on oxidative stress, hepatotoxicity, apoptosis, and inflammation in zebrafish embryos.
Co-existence of mycotoxins may pose a greater risk. It remains less known about the toxic effect of co-exposure of zearalenone (ZEA) and deoxynivalenol (DON) on aquatic life. In the present study, the toxic effects of the combine treatment of ZEA and DON on zebrafish (Danio rerio) embryos were investigated. The results showed that the combined treatment of ZEA (200, 400, 800 μg/L) and DON (4000 μg/L) did not cause apparent deaths, but induced a developmental toxicity as indicated by decreased movement times and heartbeat. At 96 h post-fertilization (hpf), co-exposure of ZEA and DON (Z400 + D4000 and Z800 + D4000 group) led to significant oxidative stress as evidenced by the increased ROS level and MDA content, as well as the changes of antioxidant enzymes (SOD, CAT and GPX) and their genes. Besides, the combined treatment of ZEA and DON triggered hepatotoxicity as shown by the changes of Fabp10a, Gclc, Gsr, Nqo1 genes, apoptosis through upregulating apoptosis-related genes (p53, Caspase-9, Caspase-3) and downregulating Bcl-2 gene, as well as inflammation by promoting the expression of IL-1β, IL-6, TNF-α, TLR4, MyD88, NF-κBp65 genes. These results indicated the co-exposure of ZEA and DON caused oxidative stress, leading to stronger potential toxic effects to zebrafish embryos than their respective single treatment. Therefore, more attention should be paid to risk management of the co-contamination of mycotoxins.


Mass spectrometry of high-mannose oligosaccharides after trifluoroacetolysis and periodate oxidation.
High-mannose oligosaccharides were treated with a mixture of trifluoroacetic acid and trifluoroacetic anhydride under conditions where reducing terminal N-acetylglucosamine residues are specifically degraded. The resulting mannose-containing products were, after further chemical modifications, analyzed by mass spectrometry in fast atom bombardment and electron ionization modes. Binding positions between monosaccharide residues were deduced from mass spectra of peracetylated compounds, which, prior to the derivatization, had been subjected to periodate oxidation and borodeuteride reduction.


Maternal diet, rather than obesity itself, has a main influence on milk triacylglycerol profile in dietary obese rats.
Triacylglycerols (TG) in milk derive from different sources, and their composition may be influenced by both maternal diet and obesity. We used two rat models to ascertain potential changes in TG composition in milk associated to maternal intake of an obesogenic diet during lactation and to distinguish them from the effects attributable to maternal adiposity. Milk samples were obtained from dams fed a cafeteria diet during lactation (CAF) and from dams made obese by cafeteria diet feeding, with dietary normalization before gestation (PCaf). Levels of specific TG species in milk collected at different time points of lactation were determined by shotgun lipidomics. CAF and PCaf dams presented a greater adiposity than their respective controls. The principal component analysis of TG peaks showed a clear separation between milk from CAF dams and milk from control and Pcaf dams, already evident at 5 days of lactation. Milk from CAF dams was enriched with TG species with greater number of carbons and double bonds and reduced in TG with lower number of carbons. TG composition of milk from Pcaf dams was similar to controls, although specific differences were observed at day 5 of lactation. Thus, the intake of a cafeteria diet during lactation, rather than maternal adiposity, alters milk composition. This effect is avoided with dietary normalization before gestation, although the remaining fat reserves may also influence TG composition at initial stages of lactation. Therefore, normalization of maternal diet prior to pregnancy should be considered as a strategy for achieving optimal milk composition.


Inhibition of H-Ras and MAPK is compensated by PKC-dependent pathways in annexin A6 expressing cells.
High-density lipoprotein (HDL)-induced activation of the Ras/MAPK pathway can be mediated by protein kinase C (PKC)-dependent and independent pathways. Although both pathways co-exist in cells, we showed that binding of HDL to scavenger receptor BI (SR-BI) in CHO cells activates Ras and MAPK in a PKC-independent manner. We have recently identified that HDL-induced activation of Ras and Raf-1 is reduced in annexin A6 expressing CHO cells (CHOanx6). In the present study we demonstrate that despite the loss of Ras and Raf-1 activity, HDL induces MAPK phosphorylation in CHOanx6 cells. Since annexin A6 is a PKCalpha-binding protein we therefore investigated the possible involvement of PKC in HDL-induced Ras and MAPK activation in CHOanx6 cells. Taken together our findings demonstrate that HDL-induced H-Ras and MAPK activation is PKC-dependent in cells expressing annexin A6 to compensate for the loss of PKC-independent activation of H-Ras and MAPK.


Nitric oxide regulates swimming in the jellyfish Aglantha digitale.
The cnidarian nervous system is considered by many to represent neuronal organization in its earliest and simplest form. Here we demonstrate, for the first time in the Cnidaria, the neuronal localization of nitric oxide synthase (NOS) in the hydromedusa Aglantha digitale (Trachylina). Expression of specific, fixative-resistant NADPH-diaphorase (NADPH-d) activity, characteristic of NOS, was observed in neurites running in the outer nerve ring at the base of the animal and in putative sensory cells in the ectoderm covering its tentacles. At both sites, diphenyleneiodonium (10(-4) M) abolished staining. Capillary electrophoresis confirmed that the NO breakdown products NO2- and NO3- were present at high levels in the tentacles, but were not detectable in NADPH-d-negative areas. The NADPH-d-reactive neurons in the tentacles send processes to regions adjacent to the inner nerve ring where swimming pacemaker cells are located. Free-moving animals and semi-intact preparations were used to test whether NO is involved in regulating the swimming program. NO (30-50 nM) and its precursor L-arginine (1 mM) stimulated swimming, and the effect was mimicked by 8-Br-cGMP (50-100 microM). The NO scavenger PTIO (10-100 microM) and a competitive inhibitor of NOS, L-nitroarginine methyl ester (L-NAME, 200 microM), significantly decreased the swimming frequency in free-moving animals, while its less-active stereoisomer D-nitroarginine methyl ester (D-NAME, 200 microM) had no such effect. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ, 5-20 microM), a selective inhibitor of soluble guanylyl cyclase, suppressed spontaneous swimming and prevented NO-induced activation of the swimming program. We suggest that an NO/cGMP signaling pathway modulates the rhythmic swimming associated with feeding in Aglantha, possibly by means of putative nitrergic sensory neurons in its tentacles.


Current status of external counterpulsation.
This article traces the development of external counterpulsation from its beginnings to the present. Initially, counterpulsation was carried out by cannulating the femoral artery. The hemodynamic goals were to reduce the afterload of the left ventricle, and to raise or augment the diastolic pressure. This gave rise to the term "counterpulsation." The intra-aortic balloon is capable of producing these salutary effects because of its proximity to the outlet of the left ventricle. The same hemodynamic effects can be obtained by external counterpulsation. However, one must produce a negative pressure during cardiac systole, and ensure that this is applied to the lower extremities. The only clinical study in which this was done was in the treatment of patients in cardiogenic shock by Soroff and colleagues. The results of the clinical studies reviewed are all suggestive of benefits derived from external counterpulsation in a variety of clinical settings. These studies suggest the following avenues for improvement in the equipment used to carry out external counterpulsation: 1. Inclusion of the vascular bed of the buttocks to be subjected to the external pressures, as advocated by Zheng. 2. Inclusion of a negative pressure blanket, as advocated by Soroff. 3. Further investigation of graded-sequential external counterpulsation, using the buttocks and negative pressure. 4. Application of external counterpulsation earlier in cardiogenic shock and for at least 4 hours in acute myocardial infarction. Our evaluation is that this method has not been studied in a way that demonstrates its full potential. We feel that it is on the threshold of being shown to be useful in all of the clinical settings reviewed, and we hope that the necessary equipment will be created to allow investigators to establish its proper place in our therapeutic armamentarium.


Variceal Bleeding: Beyond Banding.
Despite advances in the management of complications of portal hypertension, variceal bleeding continues to be associated with significant morbidity and mortality. While endoscopic variceal band ligation remains first line therapy for treating bleeding and high-risk non-bleeding esophageal varices, alternate therapies have been explored, particularly in cases of refractory bleeding. The therapies being explored include stent placement, hemostatic powder use, over-the-scope clips and others. For gastric variceal bleeding, endoscopic ultrasound-guided therapies have recently emerged as promising interventions for hemostasis. The aim of this article is to highlight these alternative therapies and their potential role in the management of gastric and esophageal variceal bleeding.


Physiologically based pharmacokinetic model for humans orally exposed to chromium.
A multi-compartment physiologically based pharmacokinetic (PBPK) model was developed to describe the behavior of Cr(III) and Cr(VI) in humans. Compartments were included for gastrointestinal lumen, oral mucosa, stomach, small intestinal tissue, blood, liver, kidney, bone, and a combined compartment for remaining tissues. As chronic exposure to high concentrations of Cr(VI) in drinking water cause small intestinal cancer in mice, the toxicokinetics of Cr(VI) in the upper gastrointestinal tract of rodents and humans are important for assessing internal tissue dose in risk assessment. Fasted human stomach fluid was collected and ex vivo Cr(VI) reduction studies were conducted and used to characterize reduction of Cr(VI) in human stomach fluid as a mixed second-order, pH-dependent process. For model development, toxicokinetic data for total chromium in human tissues and excreta were identified from the published literature. Overall, the PBPK model provides a good description of chromium toxicokinetics and is consistent with the available total chromium data from Cr(III) and Cr(VI) exposures in typical humans (i.e., model predictions are within a factor of three for approximately 86% of available data). By accounting for key species differences, sources of saturable toxicokinetics, and sources of uncertainty and variation, the rodent and human PBPK models can provide a robust characterization of toxicokinetics in the target tissue of the small intestine allowing for improved health risk assessment of human populations exposed to environmentally-relevant concentrations.


Functional oncocytoma of the kidney: evaluation by dual tracer scintigraphy.
We report a case of renal oncocytoma with a unique scintigraphic pattern. The tumor showed strong avidity for [123I]iodohippurate, but no affinity for [99mTc] glucoheptonate. We offer an explanation for such exceptional scintigraphic finding, which may potentially enable us to make a pre-operative diagnosis of renal oncocytoma in the future.


Using Resin-Based 3D Printing to Build Geometrically Accurate Proxies of Porous Sedimentary Rocks.
Three-dimensional (3D) printing is capable of transforming intricate digital models into tangible objects, allowing geoscientists to replicate the geometry of 3D pore networks of sedimentary rocks. We provide a refined method for building scalable pore-network models ("proxies") using stereolithography 3D printing that can be used in repeated flow experiments (e.g., core flooding, permeametry, porosimetry). Typically, this workflow involves two steps, model design and 3D printing. In this study, we explore how the addition of post-processing and validation can reduce uncertainty in the 3D-printed proxy accuracy (difference of proxy geometry from the digital model). Post-processing is a multi-step cleaning of porous proxies involving pressurized ethanol flushing and oven drying. Proxies are validated by: (1) helium porosimetry and (2) digital measurements of porosity from thin-section images of 3D-printed proxies. 3D printer resolution was determined by measuring the smallest open channel in 3D-printed "gap test" wafers. This resolution (400 µm) was insufficient to build porosity of Fontainebleau sandstone (∼13%) from computed tomography data at the sample's natural scale, so proxies were printed at 15-, 23-, and 30-fold magnifications to validate the workflow. Helium porosities of the 3D-printed proxies differed from digital calculations by up to 7% points. Results improved after pressurized flushing with ethanol (e.g., porosity difference reduced to ∼1% point), though uncertainties remain regarding the nature of sub-micron "artifact" pores imparted by the 3D printing process. This study shows the benefits of including post-processing and validation in any workflow to produce porous rock proxies.


Correlation of the primary Gleason pattern on prostate needle biopsy with clinico-pathological factors in Gleason 7 tumors.
To correlate the primary Gleason pattern among patients with biopsy-derived Gleason 7 tumors with the radical prostatectomy specimen Gleason grading and other clinical and pathologic outcomes. Among 474 patients who underwent radical prostatectomy for clinically localized prostate cancer between 1997-2001, 205 (43%) had Gleason 7/10 tumors on pre-operative needle biopsy. Among theses patients, 148 (72.2%) were assigned a primary Gleason 3 pattern (3+4 = 7) and 57 (27.8%) were assigned a primary Gleason 4 pattern (4+3 = 7). The two groups were compared with respect to age, serum PSA levels, Gleason grade in the radical prostatectomy specimen, pathological stage and surgical margin status. Among patients with 3+4 tumors on needle biopsy, 64% remained primary Gleason grade 3 while 35% were up-graded to a primary pattern 4 following analysis of the radical prostatectomy specimen. Patients with 4+3 tumors on needle biopsy remained primary Gleason grade 4 in 51% of patients, while 49% of patients had their tumors down-graded to a primary 3 pattern (p = 0.09). There were no differences between patients with needle biopsy 3+4 and 4+3 patterns with respect to total Gleason score in the radical prostatectomy specimen (p = 0.42), pTNM stage (p = 0.36), extra-prostatic extension (p = 0.88), surgical margin involvement (p = 0.16), and seminal vesicle invasion (p = 0.19). In contrast, the primary Gleason pattern in the radical prostatectomy specimen correlated significantly with pTNM stage (p = 0.02) and seminal vesicle invasion (p= 0.003), but not with extra-prostatic extension (p = 0.32) and surgical margin involvement (p = 0.17). Among patients with Gleason 7 adenocarcinoma of the prostate, the biopsy-derived primary Gleason pattern does not appear to correlate with important clinical and pathologic outcomes. The utility of distinguishing a primary Gleason pattern on needle biopsy among patients with Gleason 7 tumors remains unclear given the limited and conflicting literature addressing this issue.


Trends in ischaemic heart disease hospitalisation and case fatality in the Hong Kong Chinese population 2000-2009: a secondary analysis.
This study examines the recent trends in ischaemic heart disease (IHD) incidence and case fatality in Hong Kong and explores the possible risk factors. Retrospective observational study. All public hospitals in Hong Kong. Incidence rate was defined as the number of IHD inpatient episodes divided by the size of the corresponding population. Short-term and long-term case fatality rate was defined as deaths from all causes occurring within 30 and 31-365 days, respectively, divided by the number of IHD inpatient episodes among the corresponding population. Poisson and logistic regression models were used to examine the IHD incidence and short-term/long-term case fatality trends, respectively, for different age and sex groups. IHD incidence was stable in most age groups. However, the incidence in men aged 15-24, 35-44 and ≥85 years showed increasing trends, whereas the incidence in men aged 55-64 years and women aged 35-74 years showed decreasing trends. Overall, the short-term/long-term case fatality rates were unchanged over time for both sexes. Short-term case fatality showed increasing trends in women aged 65-74 and ≥85 years, while long-term case fatality in men aged 55-64 and 75-84 years and women aged ≥75 years showed increasing trends. Hong Kong trends resembled those in the USA, England and Wales, showing stable or slow decline in the IHD rates, while increasing trends were observed for some age groups, particularly young adults. Public health promotion efforts should focus on reducing cardiovascular risk factors, such as hypertension prevalence.


Trends in bacteremia in the pre- and post-highly active antiretroviral therapy era among HIV-infected children in the US Perinatal AIDS Collaborative Transmission Study (1986-2004).
HIV-infected children are at high risk for bacteremia. Highly active antiretroviral therapy has reduced rates of opportunistic infections; less is known about its effect on pediatric bacteremia rates. Thus, we sought to determine its impact on bacteremia incidence in HIV-infected children. Children born during 1986-1998 were followed until 2004 in the Perinatal AIDS Collaborative Transmission Study. We determined the pre- and post-highly active antiretroviral therapy (before and after January 1, 1997) incidence of bacteremia among HIV-infected children and characterized the CD4% temporal declines and mortality among patients with and those without incident bacteremias. Among 364 children, 68 had 118 documented bacteremias, 97 before and 21 after January 1, 1997. Streptococcus pneumoniae constituted 56 (58%) pre- and 13 (62%) post-highly active antiretroviral therapy cases. The incidence rate ratio of bacteremias comparing post- versus pre-highly active antiretroviral therapy was 0.3 overall and 0.2, 0.2, and 0.4 among children aged 0 to 24, 25 to 48, and 49 to 72 months, respectively. Kaplan-Meier analysis for time to first bacteremia in children born during the pre-highly active antiretroviral therapy compared with the post-highly active antiretroviral therapy era revealed that 69% and 94%, respectively, remained bacteremia free at a median follow-up of 6 years. The Cox proportional hazards model also showed a significant reduction of bacteremias in the post-highly active antiretroviral therapy era, even after controlling for gender and race. Among children <6 years of age, those who experienced bacteremia had faster temporal CD4% decline than those who never had bacteremia. Survival analysis revealed that HIV-infected children with bacteremia experienced higher overall mortality when controlling for gender, race, and clinic site. A significant decrease in bacteremia incidence and a prolongation in the time to first bacteremia incident were seen in the post-highly active antiretroviral therapy era. Children with a steeper decline of CD4 T cells were more likely to develop bacteremia. Children who experienced bacteremia had an associated higher mortality than their bacteremia-free counterparts.


Rev1 contributes to proper mitochondrial function via the PARP-NAD+-SIRT1-PGC1α axis.
Nucleic acids, which constitute the genetic material of all organisms, are continuously exposed to endogenous and exogenous damaging agents, representing a significant challenge to genome stability and genome integrity over the life of a cell or organism. Unrepaired DNA lesions, such as single- and double-stranded DNA breaks (SSBs and DSBs), and single-stranded gaps can block progression of the DNA replication fork, causing replicative stress and/or cell cycle arrest. However, translesion synthesis (TLS) DNA polymerases, such as Rev1, have the ability to bypass some DNA lesions, which can circumvent the process leading to replication fork arrest and minimize replicative stress. Here, we show that Rev1-deficiency in mouse embryo fibroblasts or mouse liver tissue is associated with replicative stress and mitochondrial dysfunction. In addition, Rev1-deficiency is associated with high poly(ADP) ribose polymerase 1 (PARP1) activity, low endogenous NAD+, low expression of SIRT1 and PGC1α and low adenosine monophosphate (AMP)-activated kinase (AMPK) activity. We conclude that replication stress via Rev1-deficiency contributes to metabolic stress caused by compromized mitochondrial function via the PARP-NAD+-SIRT1-PGC1α axis.


Trial of Early Antiviral Therapies during Non-hospitalized Outpatient Window (TREAT NOW) for COVID-19: a summary of the protocol and analysis plan for a decentralized randomized controlled trial.
Coronavirus disease 2019 (COVID-19) has a heterogeneous outcome in individuals from remaining asymptomatic to death. In a majority of cases, mild symptoms are present that do not require hospitalization and can be successfully treated in the outpatient setting, though symptoms may persist for a long duration. We hypothesize that drugs suitable for decentralized study in outpatients will have efficacy among infected outpatients METHODS: The TREAT NOW platform is designed to accommodate testing multiple agents with the ability to incorporate new agents in the future. TREAT NOW is an adaptive, blinded, multi-center, placebo-controlled superiority randomized clinical trial which started with two active therapies (hydroxychloroquine and lopinavir/ritonavir) and placebo, with the hydroxychloroquine arm dropped shortly after enrollment began due to external evidence. Each arm has a target enrollment of 300 participants who will be randomly assigned in an equal allocation to receive either an active therapy or placebo twice daily for 14 days with daily electronic surveys collected over days 1 through 16 and on day 29 to evaluate symptoms and a modified COVID-19 ordinal outcome scale. Participants are enrolled remotely by telephone and consented with a digital interface, study drug is overnight mailed to study participants, and data collection occurs electronically without in-person interactions. If effective treatments for COVID-19 can be identified for individuals in the outpatient setting before they advance to severe disease, it will prevent progression to more severe disease, reduce the need for hospitalization, and shorten the duration of symptoms. The novel decentralized, "no touch" approach used by the TREAT NOW platform has distinction advantages over traditional in-person trials to reach broader populations and perform study procedures in a pragmatic yet rigorous manner. ClinicalTrials.gov NCT04372628. Registered on April 30, 2020. First posted on May 4, 2020.


Evaluation of a new multi-epitope sequence of eight known Leishmania infantum antigens for HVL diagnosis by ELISA and Western blot.
Leishmaniasis, known as a disease with high prevalence proportion throughout the world, is caused by protozoan parasites. Visceral leishmaniasis is the most severe form of this condition reported sporadically from all regions in Iran. Between different diagnostic tests, serodiagnosis of this infection is of utmost importance in both humans and dogs. Although rK39 ELISA test has been extensively validated in endemic areas, there are currently challenges regarding a more appropriate serodiagnostic test. A novel multi-epitope construct was designed consisting of highexposedB cell epitopes using eight important antigens of Leishmania infantum (Gp63, KMP-11, HSP70, CPA, H2A, H3, LACK and TRYP). Our artificial sequence, a Multi-epitope Recombinant Protein (MRP), was consequently produced and purified. Then, immunoreactivity was investigated by ELISA test and western blotting as well. In the present study, the cutoff value (1.052) for the new MRP-ELISA was determined by receiver operator characteristic (ROC) curve analysis using 35 known positive and 20 known negative HVL sera previously tested for antibodies to L. infantum by DAT, showing a sensitivity of 93.1% and a specificity of 77.4%. The blotting test also showed a favorable band to detect visceral leishmaniasis. According to the results, this new antigen had acceptable potential in detecting VL positive cases once western blotting was utilized, but the ELISA test did not proceed as expected for detecting true negative cases, probably due to some optimization issues.The present study is a promising start.


Protection provided by exogenous DNA ligase in G0 human lymphocytes treated with restriction enzyme MspI or bleomycin as shown by the comet assay.
DNA double-strand breaks (DSB) may arise either spontaneously during cellular processes or as a result of exposure to DNA-damaging agents such as ionizing radiation, or radiomimetic agents such as restriction endonucleases or bleomycin. It is widely accepted that nonrepaired or misrepaired DSB are the main lesions leading to the production of chromosomal aberrations, mutagenesis, oncogenic transformation, and cell killing. Studies focusing on this relationship, as well as the possible modulation of DNA repair mechanisms, are currently of major interest. A wide variety of test systems are available to study DNA damage. In the last few years, single-cell gel electrophoresis, commonly known as "comet assay," has been considered a rapid, sensitive, and visual method for quantifying DNA strand breaks and alkali-labile damage in individual cells. In this study, making use of the comet assay, we tried to find out if under conditions that maintain chromatin structure the DNA ligase from T4 phage is able to facilitate the rejoining of strand breaks with different end structures, induced by the restriction endonuclease MspI or bleomycin in living human lymphocytes in a nonproliferating state. T4 DNA ligase, as well as the restriction endonuclease or bleomycin, were introduced together by electroporation into human lymphocytes. Our results support the idea that it is possible to modulate the DSB-rejoining of different DNA strand-breaking agents by exogenous T4 DNA ligase.


Semi-automated computed tomography Volumetry can predict hemihepatectomy specimens' volumes in patients with hepatic malignancy.
One of the major causes of perioperative mortality of patients undergoing major hepatic resections is post-hepatectomy liver failure (PHLF). For preoperative appraisal of the risk of PHLF it is important to accurately predict resectate volume and future liver remnant volume (FLRV). The objective of our study is to prospectively evaluate the accuracy of hemihepatectomy resectate volumes that are determined by computed tomography volumetry (CTV) when compared with intraoperatively measured volumes and weights as gold standard in patients undergoing hemihepatectomy. Twenty four patients (13 women, 11 men) scheduled for hemihepatectomy due to histologically proven primary or secondary hepatic malignancies were included in our study. CTV was performed using a semi-automated module (S, hereinafter) (syngo.CT Liver Analysis VA30, Siemens Healthcare, Germany). Conversion factors between CT volumes on the one side and intraoperative volumes and weights on the other side were calculated using the method of least squares. Absolute and relative disagreements between CT volumes and intraoperative volumes were determined. A conversion factor of c = 0.906 most precisely predicted intraoperative volumes of exsanguinated hemihepatectomy specimens from CT volumes in all patients with mean absolute and relative disagreements between CT volumes and intraoperative volumes of 57 ml and 6.3%. The use of operation-specific conversion factors yielded even better results. CTV performed with S accurately predicts intraoperative volumes of hemihepatectomy specimens when applying conversion factors which compensate for exsanguination. This allows to precisely estimate the FLRV and thus minimize the risk of PHLF in patients undergoing major hepatic resections.


Using bayesian model to estimate the cost of traffic injuries in Iran in 2013.
A significant social and economic burden inflicts by road traffic injuries (RTIs). We aimed to use Bayesian model, to present the precise method, and to estimate the cost of RTIs in Iran in 2013. In a cross-sectional study on costs resulting from traffic injuries, 846 people per road user were randomly selected and investigated during 3 months (1st September-1st December) in 2013. The research questionnaire was prepared based on the standard for willingness to pay (WTP) method considering perceived risks, especially in Iran. Data were collected along with four scenarios for occupants, pedestrians, vehicle drivers, and motorcyclists. Inclusion criterion was having at least high school education and being in the age range of 18-65 years old; risk perception was an important factor to the study and measured by visual tool. Samples who did not have risk perception were excluded from the study. Main outcome measure was cost estimation of traffic injuries using WTP method. Mean WTP was 2,612,050 internal rate of return (IRR) among these road users. Statistical value of life was estimated according to 20,408 death cases 402,314,106,073,648 IRR, equivalent to 13,410,470,202$ based on the dollar free market rate of 30,000 IRR (purchase power parity). In sum, injury and death cases came to 1,171,450,232,238,648 IRR equivalents to 39,048,341,074$. Moreover, in 2013, costs of traffic accident constituted 6.46% of gross national income, which was 604,300,000,000$. WTP had a significant relationship with age, middle and high income, daily payment to injury reduction, more payment to time reduction, trip mileage, private cars drivers, bus, minibus vehicles, and occupants (P < 0.01). Costs of traffic injuries included noticeable portion of gross national income. If policy-making and resource allocation are made based on the scientific pieces of evidence, an enormous amount of capital can be saved through reducing death and injury rates.


One-stage microvascular mandible reconstruction and alloplastic TMJ prosthesis.
Severely deformed or absent temporomandibular joints (TMJ) benefit from total alloplastic joint replacement and large mandibular defects from revascularized free tissue transfer for reconstruction. However no cases of their combined one-stage placement with outcomes can be found in the literature. We present two cases with different indications and reconstruction. The first patient required mandibular body and ascending ramus reconstruction after previous sarcoma resection. This was with a condyle-bearing reconstruction plate which resulted in significant dysfunction, leaving the patient unable to open her mouth. A one-stage vascularized iliac crest free flap and alloplastic TMJ prosthesis was used to reconstruct the mandible. Subsequently, metal removal, soft tissue augmentation by lipotransfer and dental implant placement were performed. At 63 months follow-up patient was pain-free, with mouth opening, protrusion and lateral excursion back to normal. The second patient required mandibular body, ascending ramus and joint reconstruction, performed by transoral vascularized fibula free flap with temporal vessel anastomosis. The traumatic deep bite and posterior facial height were corrected, additional submandibular scars avoided by transoral placement of the fibula transplant and a miniaturized TMJ prosthesis along with the vascularised free flap with 28 months follow-up. A miniaturized TMJ prosthesis may become placed transorally for reconstruction of the TMJ, together with a vascularized free flap for mandibular reconstruction and promises good long-term stability with normal function above all for protrusion and lateral excursion.


Increasing incidence and continued dismal outcome of primary central nervous system lymphoma in Norway 1989-2003 : time trends in a 15-year national survey.
The incidence of primary central nervous system lymphoma (PCNSL) appears to be increasing in some countries, whereas it is stable in others. Many reports the last decades have suggested that there have been improvements in the treatment of PCNSL. The objective of this study was to analyze time trends in the incidence, clinical features, histologic diagnosis, treatment, and outcome of nonacquired immunodeficiency syndrome (non-AIDS) PCNSL in Norway from 1989 to 2003. Patients were identified by a chart review of all patients who had a recorded diagnosis of PCNSL from 1989 to 2003 in The Norwegian Cancer Registry. The histologic and cytologic material from each patient was re-examined by pathologists. Time trends were analyzed according to year of diagnosis grouped into 3 5-year periods: 1989-1993, 1994-1998, and 1999-2003. There were 98 patients who had confirmed, newly diagnosed non-AIDS PCNSL in Norway from 1989 to 2003. The incidence rate increased during the consecutive 5-year periods from 0.89 per million during 1989 to 1993, to 1.74 per million during 1994 to 1998, and to 1.82 per million during 1999 to 2003 (P = .013). Diagnostic delay and overall survival did not improve with time. Survival decreased from 1999 to 2003 compared with survival from 1994 to 1998, which was explained in part by reduced performance status and fewer patients receiving combined chemotherapy and radiotherapy during 1999 to 2003. In multivariate analysis, age </=50 years, a good performance status, and active treatment (especially combined chemotherapy and radiotherapy) significantly improved survival. The incidence of PCNSL is increasing in Norway. Despite diagnostic and therapeutic advances over the last decades, neither a reduction in diagnostic delay nor any improvement in overall survival with time was observed. The search for improved understanding of etiology and treatment should be intensified.


Tartrazine and prostaglandin-system.
Tartrazine, a dye largely employed for colouring foods, drinks, drugs and cosmetics, induces in some aspirin-sensitive subjects a bronchoconstriction similar to that caused by aspirin and other nonsteroidal anti-inflammatory drugs. The interference of tartrazine on prostaglandin system of guinea pig lungs perfused with arachidonic acid has been studied. This compound is able to inhibit in the same manner the formation both of prostaglandins and of thromboxane A2 acting at the cyclooxygenase level as has already been demonstrated for aspirin and indomethacin. Preliminary experiments on aspirin asthmatic patients treated or not with tartrazine are discussed.


In vitro inhibition of human cytochrome P450 enzymes by licoisoflavone B from Glycyrrhiza uralensis Fisch. ex DC.
Glycyrrhiza uralensis Fisch. ex DC, one of the 3 pharmacopeial species of licorice and widely used in dietary supplements, can inhibit certain cytochrome P450 (CYP) enzymes. Thereby, G. uralensis preparations have the potential to cause pharmacokinetic drug interactions when consumed along with prescription medicines. One compound (1.34 mg dry weight) responsible for inhibiting CYP2B6, CYP2C8, and CYP2C9 was isolated using bioactivity-guided fractionation from 250 g dried roots, stolons, and rhizomes. The enzyme kinetics and mechanisms of inhibition were determined using human liver microsomes, recombinant enzymes, and UHPLC-MS/MS-based assays. Identified as licoisoflavone B, this compound displayed reversible inhibition of CYP2C8 with an IC50 value of 7.4 ± 1.1 µM and reversible inhibition of CYP2C9 with an IC50 value of 4.9 ± 0.4 µM. The enzyme kinetics indicated that the mechanism of inhibition was competitive for recombinant CYP2C8, with a Ki value of 7.0 ± 0.7 μM, and mixed-type inhibition for recombinant CYP2C9, with a Ki value of 1.2 ± 0.2 μM. Licoisoflavone B moderately inhibited CYP2B6 through a combination of irreversible and reversible mechanisms with an IC50 value of 16.0 ± 3.9 µM.


Ribonucleotide reductase activity and growth of glutathione-depleted mouse leukemia L1210 cells in vitro.
L1210 cells treated with L-buthionine-(S/R)-sulfoximine (BSO) had glutathione (GSH) and non-protein thiol levels only 15% that of control. These GSH-depleted cells grew as well as the control L1210 cells and there was no decrease in ribonucleotide reductase activity in situ as measured by the conversion of [14C]cytidine to deoxytidine nucleotides and incorporation into DNA. Further, when these BSO-stressed cells were treated with hydroxyurea or IMPY, there was no potentiation of the inhibition caused by hydroxyurea or IMPY alone. These data indicate that the glutathione/glutaredoxin system of ribonucleotide reductase is not the sole carrier of reducing equivalents from NADPH for the reduction of the 2'-position of the corresponding ribonucleoside 5'-diphosphate; and that glutathione is not critical in regenerating the tyrosyl free-radical on the M2 subunit which is destroyed by the hydroxyurea or 2,3-dihydro-1H-pyrazolo-[2,3-alpha]imidazole (IMPY) treatment.


Evaluation of malaria clinics in Maesot, Thailand: use of serology to assess coverage.
The clinics of the anti-malaria programme in Thailand serve an increasingly important role in the strategy for control of malaria within a context of multi-drug resistant falciparum malaria. Figures from clinics in Maesot District show a predominance of young males among positive cases treated (56% of all cases). In contrast, sero-epidemiological findings from a random sample of over 500 villagers in the area show similar exposure rates among males and females of equal age. There were no statistically significant differences between males and females 0-15 and 16-30 years old in percentages positive by indirect fluorescent antibody tests or enzyme-linked immunosorbent assays (ELISA), mean level of ELISA positivity, or rate of sero-conversion. Differences in level of positivity did occur between males and females over 30. An index constructed from the serological findings indicated under-representation of children and women of all ages in clinics but suggested that coverage of children could be improved by the provision of a community-based, fixed-schedule mobile clinic.


Defective MNiFeO (M = Cu, Zn, Co, Mn) NRs derived from cation-exchanged Fe2Ni-MOFs for catalytic nitroarene hydrogenation.
Catalytic hydrogenation reduction provides a potential route to detoxify nitro compounds. Spinel NiFe2O4 (NiFeO) serves as a natural abundance and low-cost catalyst but suffers from low catalytic activity due to poor redox nature and limited active sites. Herein, defective MNiFeO (M = Cu, Zn, Co, Mn) nanorods (NRs) were synthesized by pyrolysis of cation-exchanged Fe2Ni-MOFs. The introduction of M modified the electronic structure of NiFeO and thus accelerated the electron transfer and proton transport in the hydrogenation of nitro aromatics. CuNiFeO exhibited the highest catalytic activity with a turn frequency of 2.89 × 105h-1, giving 308- and 422-fold larger than that of NiFeO-Vo and NiFeO in the reduction of 4-nitrophenol (4-NP). CuNiFeO NRs also presented exceptional performances in the reduction of the other nitroarenes. The remarkable improvement in activity of mesoporous CuNiFeO may be attributed to its ternary composites, and increased oxygen vacancies. This work not only provides a simple route to constructing mesoporous MNiFeO NRs with oxygen vacancies, but also further discusses the effects of metal component and oxygen defects in the catalytic reduction of 4-NP.


Principles of rat subcortical forebrain organization: a study using histological techniques and multiple fluorescence labeling.
In the present study, we introduce new views on neuro- and chemoarchitectonics of the rat forebrain subcortex deduced from traditional and current concepts of anatomical organization and from our own results. It is based on double and triple immunofluorescence of markers for transmitter-related enzymes, calcium-binding proteins, receptor proteins, myelin basic protein (MBP) and neuropeptides, and on histological cell/myelin stains. The main findings can be summarized as follows: (i) the dorsal striatum of rat and other myomorph rodents reveals a small caudate equivalent homotopic to the caudate nucleus (C) of other mammals, and a large putamen (Pu). (ii) Shell and core can be distinguished also in the 'rostral pole' of nucleus accumbens (ACC) with the calretinin/calbindin and neuropeptide Y (NPY) immunostaining. The shell reveals characteristics of a genuine striatal but not of an extended amygdala (EA) subunit. (iii) EA and lateral septum show striking similarities in structure and fiber connections and may therefore represent a separate parastriatal complex. (iv) The meandering dense layer (DL) of olfactory tubercle (OT) forms longitudinal gyrus- and sulcus-like structures converging in its rostral pole. (v) The core regions of the islands of Calleja that border the ventral pallidum (VP) sharing some of its features are invaded by myelinated fibers of the medial forebrain bundle (MFB). The island of Calleja magna is also apposed to an inconspicuous, slender dorsal appendage of VP. (vi) The VP is composed of a large dorsal reticulated part traversed by the myelinated GABAergic parvalbumin-immunoreactive axons of the MFB and a slender ventral non-reticulate part close to the islands of Calleja. (vii) Considering their close association to the limbic system, ventral striatum (VS) and VP may represent the oldest part of basal ganglia, whereas dorsal striatopallidal subunits were progressively developed in parallel to the growing neocortical influence on motor behavior.


A self-powered bidirectional partition microfluidic chip with embedded microwells for highly sensitive detection of EGFR mutations in plasma of non-small cell lung cancer patients.
Circulating tumor DNA (ctDNA) is a promising biomarker for tumor genotyping and therapy monitoring. Herein, we developed a digital PCR chip with embedded microwell and bidirectional partition network for highly sensitive ctDNA analysis. The embedded microwell contributes to increasing microreaction density (up to 7000 microwells/cm2) and reducing evaporation during amplification. The bidirectional partition network can achieve fast and random distribution of targets, ensuring the precise quantification of nucleic acid. We used plasmids, artificial samples and 32 clinical blood samples from non-small cell lung cancer patients to test the performance of this platform. The results demonstrated that our chip has not only comparable quantification performance to commercial counterpart but also the ability to detect EGFR mutations with as low as 0.01% mutation rate and 20 alter molecules in 27 ng genomic DNA. The identification of EGFR mutations in plasma using developed chip exhibited 85.71% sensitivity and 94.44% specificity for L858R mutation and 100% sensitivity and 86.96% specificity for T790 M mutation. Moreover, the monitoring of mutant allele in plasma was accomplished in this work. In conclusion, the developed chip has a potential in lung tumor genotyping and therapy monitoring for precision medicine, even other tumors.


Which imaging modality in cochlear implant candidates?
There is no guideline or consensus on preoperative radiologic imaging modality despite the fact that it has a vital importance in appropriate candidacy selection of cochlear implantation. We aimed to find out the role of high-resolution computed tomography (HRCT) and magnetic resonance imaging (MRI) on surgical planning, intraoperative technique in cochlear implant candidates. The clinical charts, imagings, and operative reports of patients who underwent cochlear implant surgery at a tertiary institution were retrospectively examined. 611 patients (503 children and 108 adult) were enrolled into the study. We found 11 different pathologies in MRI which could not be seen in HRCT. However, we decided the side of surgery according to MRI in only three of them in which the pathology was cochlear nerve hypoplasia. Two patients with cochlear nerve hypoplasia were children with prelingual deafness and one was adult with perilingual deafness. Moreover, we changed the surgical planning of side according to both imaging modalities in nine patients. Seven of them were children and two were adult. One of these adults had cochlear anomaly, and another had bilateral temporal bone fracture. We suggest both imaging modalities in pediatric candidates. However, in adults, we think that superiority of either imaging modalities is still contradictive. We had only three adult patients and the decision of the side of surgery was made according to MRI in one of them and to both imaging modalities in the other two adults.


Variability of blood lead levels in an urban population in relation to drinking and smoking habits.
The influence of drinking and smoking habits on blood lead levels of an urban population was assessed in this study. Lead concentrations were determined in 287 blood samples collected from non-occupationally lead exposed men living in Barcelona, Spain. Blood lead levels significantly increased with the consumption of wine. Although not statistically significant, a similar tendency was also observed for beer drinkers, but not for the consumers of spirits. Slight increases in the blood lead concentrations of smokers were also found. However, because of the significant correlation between heavy smoking and drinking observed in this study, alcohol consumption would probably be responsible for these increases. Wine consumption was the most influential variable (7.72%, P < 0.001) on the blood lead levels of the population examined.


Fluorous-Phase Ion-Selective pH Electrodes: Electrode Body and Ionophore Optimization for Measurements in the Physiological pH Range.
Because of their low polarity and polarizability, fluorous sensing membranes are both hydrophobic and lipophobic and exhibit very high ion selectivities. Here, we report on a new fluorous-membrane ion-selective electrode (ISE) with a wide sensing range centered around physiologically relevant pH values. The fluorophilic tris[perfluoro(octyl)butyl]amine (N[(CH2)4Rf8]3) was synthesized and tested as a new H+ ionophore using a redesigned electrode body that provides excellent mechanical sealing and much improved measurement reliability. In a challenging 1 M KCl background, these fluorous-phase ISEs exhibit a sensing range from pH 2.2 to 11.2, which is one of the widest working ranges reported to date for ionophore-based H+ ISEs. High selectivities against common interfering ions such as K+, Na+, and Ca2+ were determined (selectivity coefficients: logK H, K pot = - 11.6; logK H, Na pot = - 12.4; logK H, Ca pot < - 10.2). The use of the N[(CH2)4Rf8]3 ionophore with its -(CH2)4- spacers separating the amino group from the strongly electron-withdrawing perfluorooctyl groups improved the potentiometric selectivity as compared to the less basic tris[perfluoro(octyl)propyl]amine ionophore. The use of N[(CH2)4Rf8]3 also made the ISE less prone to counter anion failure (i.e., Donnan failure) at low pH than the use of tris[perfluoro(octyl)pentyl]amine with its longer -(CH2)5- spacers, which more effectively shield the amino center from the perfluorooctyl groups. In addition, we exposed both conventional plasticized PVC-phase pH ISEs and fluorous-phase pH ISEs to 10% serum for 5 days. Results show that the PVC-phase ISEs lost selectivity while their fluorous-phase counterparts did not.


cGMP stimulates endoplasmic reticulum Ca(2+)-ATPase in vascular endothelial cells.
Calcium is a crucial regulator of many physiological processes such as cell growth, division, differentiation, cell death and apoptosis. In this study, we examined the effect of cGMP on agonist-induced [Ca(2+)](i) transient in isolated rat aortic endothelial cells. 100 microM ATP was applied to the cells bathed in a Ca(2+)-free physiological solution to induce a [Ca(2+)](i) transient that was caused by Ca(2+) release from intracellular stores. cGMP, which was applied after [Ca(2+)](i) reached its peak level, accelerated the falling phase of [Ca(2+)](i) transient. Pre-treatment of the cells with CPA abolished the accelerating effect of cGMP on the falling phase of [Ca(2+)](i) transient. The effect of cGMP was reversed by KT5823, a highly specific inhibitor of protein kinase G. Taken together, these data suggest that cGMP may reduce [Ca(2+)](i) level by promoting Ca(2+) uptake through sarcoplasmic/endoplasmic reticulum ATPase and that the effect of cGMP may be mediated by protein kinase G.


Orexin receptors in the CA1 region of hippocampus modulate the stress-induced antinociceptive responses in an animal model of persistent inflammatory pain.
Stress activates multiple neural pathways and neurotransmitters that often suppress pain perception, the phenomenon called stress-induced analgesia (SIA). Orexin neurons from the lateral hypothalamus project to entire brain structures such as the hippocampus. The present study examined this hypothesis that orexinergic receptors in the CA1 region of the hippocampus may play a modulatory role in the development of SIA in formalin test as an animal model of persistent inflammatory pain. One hundred-two adult male Wistar rats were administered with intra-CA1 orexin-1 receptor (OX1r) antagonist, SB334867, at the doses of 3, 10, 30, and 100 nmol or TCS OX2 29 as orexin-2 receptor (OX2r) antagonist at the doses of 1, 3, 10, and 30 nmol. Five min later, rats were exposed to forced swim stress (FSS) for a 6-min period. Then, pain-related behaviors induced by formalin injection were measured at the 5-min blocks during a 60-min period of formalin test. The current study indicated that solely stress exposure elicits antinociception in the early and late phases of the formalin test. The FSS-induced analgesia was prevented by intra-CA1 administration of SB334867 or TCS OX2 29 during either phase of the formalin test. Moreover, the contribution of the OX2r in the mediation of analgesic effect of stress was more prominent than that of the OX1r during both phases of the formalin test. It is suggested that OX1r and OX2r in the CA1 region of the hippocampus are involved in stress-induced analgesia in the animal model of persistent inflammatory pain.


[Bilateral and monolateral dolichoarteriopathies (Kinking, Coiling, Tortuosity) of the carotid arteries and atherosclerotic disease. An ultrasonographic study].
The aim of this study, performed in outpatients, is to evaluate, using echo color Doppler examination, atherosclerotic lesions of extracranial carotid arteries in subjects with bilateral, and monolateral dolichoarteriopathies. A total of 316 subjects, 77 with bilateral and 239 with monolateral dolichoarteriopathies, were studied. Patients were divided into: 79 subjects without cardiovascular risk factors and 237 subjects with cardiovascular risk factors (98 with arterial hypertension alone, 85 with arterial hypertension associated with other cardiovascular risk factors, 54 with hypercholesterolemia, diabetes, cigarette smoking, alone or associated). Myointimal thickness, as a mean of 5 measurements at common carotid level, 2-3 cm from flow-divider and plaques as a focal thickness =/> 2 mm, were evaluated. In 77 patients with bilateral dolichoarteriopathies, compared to 239 patients with monolateral dolichoarteriopathies, there were no differences in myointimal thickening, and in the prevalence of carotid plaques; no difference was observed between sexes. Both in subjects with and without cardiovascular risk factors myointimal thickness and carotid plaques didn't show any significant differences in bilateral compared with monolateral dolichoarteriopathies. In conclusion, bilateral, compared with monolateral, dolichoarteriopathies of the carotids, do not seem to be a consequence of atherosclerotic lesions.


Protocol for fine casting, imaging, and analysis of murine vascular networks with VALID.
The majority of fluorescent vessel labeling techniques currently available are limited by their expense, incomplete labeling, or complexity. Here, we present VALID (vessel labeling via gelatin-based lipophilic dye solution)-a protocol for complete labeling of different vascular networks. We describe steps for preparing different dye hydrogels, murine vascular casting and tissue harvesting, immunolabeling, tissue clearing, and imaging, as well as detailed analysis of the vascular networks. This protocol is helpful for evaluating vascular lesions in studying different vessel-associated diseases. For complete details on the use and execution of this protocol, please refer to Zhu et al.1.


Assessing trainee's performance using reported observations: Perceptions of nurse meta-assessors.
Educational and health care organizations who prepare meta-assessors to fulfill their role in the assessment of trainees' performance based on reported observations have little literature to rely on. While the assessment of trainees' performance based on reported observations has been operationalized, we have yet to understand the elements that can affect its quality fully. Closing this gap in the literature will provide valuable insight that could inform the implementation and quality monitoring of the assessment of trainees' performance based on reported observations. The purpose of this study was to explore the elements to consider in the assessment of trainees' performance based on reported observations from the perspectives of meta-assessors. Design, Settings, Participants, data collection and analysis. The authors adopted Sandelowski's qualitative descriptive approach to interview nurse meta-assessors from two nursing programs. A semi-structured interview guide was used to document the elements to consider in the assessment of nursing trainees' performance based on reported observations, and a survey was used to collect sociodemographic data. The authors conducted a thematic analysis of the interview transcripts. Thirteen meta-assessors participated in the study. Three core themes were identified: (1) meta-assessors' appropriation of their perceived assessment roles and activities, (2) team climate of information sharing, and (3) challenges associated with the assessment of trainees' performance based on reported observations. Each theme is comprised of several sub themes. To optimize the quality of the assessment of the trainee's performance based on reported observations and ratings, HPE programs might consider how to clarify better the meta-assessor's roles and activities, as well as how interventions could be created to promote a climate of information sharing and to address the challenges identified. This work will guide educational and health care organizations for better preparation and support for meta-assessors and preceptors.


Health Care Provider Adoption of eHealth: Systematic Literature Review.
eHealth is an application of information and communication technologies across the whole range of functions that affect health. The benefits of eHealth (eg, improvement of health care operational efficiency and quality of patient care) have previously been documented in the literature. Health care providers (eg, medical doctors) are the key driving force in pushing eHealth initiatives. Without their acceptance and actual use, those eHealth benefits would be unlikely to be reaped. To identify and synthesize influential factors to health care providers' acceptance of various eHealth systems. This systematic literature review was conducted in four steps. The first two steps facilitated the location and identification of relevant articles. The third step extracted key information from those articles including the studies' characteristics and results. In the last step, identified factors were analyzed and grouped in accordance with the Unified Theory of Acceptance and Use of Technology (UTAUT). This study included 93 papers that have studied health care providers' acceptance of eHealth. From these papers, 40 factors were identified and grouped into 7 clusters: (1) health care provider characteristics, (2) medical practice characteristics, (3) voluntariness of use, (4) performance expectancy, (5) effort expectancy, (6) social influence, and (7) facilitating or inhibiting conditions. The grouping results demonstrated that the UTAUT model is useful for organizing the literature but has its limitations. Due to the complex contextual dynamics of health care settings, our work suggested that there would be potential to extend theories on information technology adoption, which is of great benefit to readers interested in learning more on the topic. Practically, these findings may help health care decision makers proactively introduce interventions to encourage acceptance of eHealth and may also assist health policy makers refine relevant policies to promote the eHealth innovation.


Postdeployment threat-related attention bias interacts with combat exposure to account for PTSD and anxiety symptoms in soldiers.
Recent studies suggest that assessment of threat-related attention bias may be useful in identifying soldiers at risk for clinical symptoms. The present study assessed the degree to which soldiers experienced combat events and showed attentional threat avoidance affected their reported levels of post-traumatic stress disorder (PTSD) and anxiety symptoms. Four months after a combat deployment to Iraq, 63 US soldiers completed a survey assessing combat exposures and clinical symptoms as well as a dot-probe task assessing threat-related attention bias. Significant three-way interactions regressing threat reaction times (RTs), neutral RTs, and combat exposure on PTSD and anxiety symptoms were observed. Specifically, soldiers with high levels of combat exposure, who were more likely to demonstrate attentional bias away from threat, were also more symptomatic. These results demonstrate the potential of threat-related attention bias as a behavioral marker of PTSD and anxiety symptoms in a high-risk military occupational context.


Accumulation of carotenoids in structural and regulatory mutants of the bacterium Myxococcus xanthus.
Accumulation of carotenoids in Myxococcus xanthus is absolutely dependent on illumination with blue light. We report the analysis of the carotenoids of dark- and light-grown cultures of the wild type and several previously characterized mutants. A carR mutant produces the same carotenoids in the dark as the wild type grown in the light. This agrees with previous evidence indicating that the carR gene codes for a general negative regulator of the system. A cis-dominant mutation in the gene carA causes constitutive expression of the light-inducible gene carB, which is linked to carA. In the dark, the carA mutant produces high levels of phytoene, the first C40 colourless carotenoid precursor; in the light, it produces the same carotenoids as the wild type. Since a mutation in carB blocks accumulation of phytoene, we propose that carB, and probably other linked genes also controlled by carA, code for enzymes involved in the synthesis of phytoene. This is virtually the only carotene accumulated by strains mutated in the gene carC, which is unlinked to the others. Thus carC codes for phytoene dehydrogenase, the enzyme that converts phytoene into coloured carotenoids. The results presented here also provide evidence for control of carotenogenesis by an endproduct that is independent of the blue light effect.


Ferroptosis-related gene signature correlates with the tumor immune features and predicts the prognosis of glioma patients.
Glioma is a malignant intracranial tumor and the most fatal cancer. The role of ferroptosis in the clinical progression of gliomas is unclear. Univariate and least absolute shrinkage and selection operator (Lasso) Cox regression methods were used to develop a ferroptosis-related signature (FRSig) using a cohort of glioma patients from the Chinese Glioma Genome Atlas (CGGA), and was validated using an independent cohort of glioma patients from The Cancer Genome Atlas (TCGA). A single-sample gene set enrichment analysis (ssGSEA) was used to calculate levels of the immune infiltration. Multivariate Cox regression was used to determine the independent prognostic role of clinicopathological factors and to establish a nomogram model for clinical application. We analyzed the correlations between the clinicopathological features and ferroptosis-related gene (FRG) expression and established an FRSig to calculate the risk score for individual glioma patients. Patients were stratified into two subgroups with distinct clinical outcomes. Immune cell infiltration in the glioma microenvironment and immune-related indexes were identified that significantly correlated with the FRSig, the tumor mutation burden (TMB), copy number alteration (CNA), and immune checkpoint expression was also significantly positively correlated with the FRSig score. Ultimately, an FRSig-based nomogram model was constructed using the independent prognostic factors age, World Health Organization (WHO) grade, and FRSig score. We established the FRSig to assess the prognosis of glioma patients. The FRSig also represented the glioma microenvironment status. Our FRSig will contribute to improve patient management and individualized therapy by offering a molecular biomarker signature for precise treatment.


Mechanisms Underlying Astrocyte Endfeet Swelling in Stroke.
Astrocyte endfeet envelop the cerebral capillaries that form the blood-brain barrier. Swelling of these endfeet occurs early in cerebral ischemia. It is generally hypothesized that such swelling occurs as the result of factors released from parenchymal brain cells during an ischemic stroke (e.g., K(+) and L-glutamate). In this review of mechanisms that can elicit astrocyte swelling in ischemic stroke, we hypothesize that, instead or in addition, such swelling may be a response to blood-brain barrier dysfunction. Astrocyte endfeet swelling may help form a cuff around a damaged vessel that limits the egress of plasma constituents and blood (hemorrhage) into brain.


Effects of 1Hz repetitive transcranial magnetic stimulation on autism with intellectual disability: A pilot study.
To explore whether 1 Hz repetitive transcranial magnetic stimulation (rTMS) has positive effects on brain activity and behavior of autistic children with intellectual disability. 32 autistic children with intellectual disability (26 boys and 6 girls) were recruited to participate in this feasibility study. The autistic children were divided randomly and equally into an experimental group and a control group. 16 children (three girls and 13 boys; mean ± SD age: 7.8 ± 2.1 years) who received rTMS treatment twice a week were served as the experimental group, while 16 children (three girls and 13 boys; mean ± SD age: 7.2 ± 1.6 years) with sham stimulation were considered as the control group. Recurrence quantification analysis (RQA) was employed to quantify the nonlinear features of electroencephalogram (EEG) signals recorded during the resting state. Three RQA measures, including recursive rate (RR), deterministic (DET) and mean diagonal length (L) were extracted from the EEG signals to characterize the deterministic features of cortical activity. Significant differences in RR and DET were observed between the experimental group and the control group. We also found discernible discrepancies in the Autism Behavior Checklist (ABC) score pre- and post-rTMS for the experimental group. 1 Hz repetitive transcranial magnetic stimulation (rTMS) could positively influence brain activity and behavior of autistic children with intellectual disability.


Antidotal treatment of botulism in rats by continuous infusion with 3,4-diaminopyridine.
Botulinum neurotoxins (BoNTs) are highly potent, select agent toxins that inhibit neurotransmitter release at motor nerve terminals, causing muscle paralysis and death by asphyxiation. Other than post-exposure prophylaxis with antitoxin, the only treatment option for symptomatic botulism is intubation and supportive care until recovery, which can require weeks or longer. In previous studies, we reported the FDA-approved drug 3,4-diaminopyridine (3,4-DAP) reverses early botulism symptoms and prolongs survival in lethally intoxicated mice. However, the symptomatic benefits of 3,4-DAP are limited by its rapid clearance. Here we investigated whether 3,4-DAP could sustain symptomatic benefits throughout the full course of respiratory paralysis in lethally intoxicated rats. First, we confirmed serial injections of 3,4-DAP stabilized toxic signs and prolonged survival in rats challenged with 2.5 LD50 BoNT/A. Rebound of toxic signs and death occurred within hours after the final 3,4-DAP treatment, consistent with the short half-life of 3,4-DAP in rats. Based on these data, we next investigated whether the therapeutic benefits of 3,4-DAP could be sustained throughout the course of botulism by continuous infusion. To ensure administration of 3,4-DAP at clinically relevant doses, three infusion dose rates (0.5, 1.0 and 1.5 mg/kg∙h) were identified that produced steady-state serum levels of 3,4-DAP consistent with clinical dosing. We then compared dose-dependent effects of 3,4-DAP on toxic signs and survival in rats intoxicated with 2.5 LD50 BoNT/A. In contrast to saline vehicle, which resulted in 100% mortality, infusion of 3,4-DAP at ≥ 1.0 mg/kg∙h from 1 to 14 d after intoxication produced 94.4% survival and full resolution of toxic signs, without rebound of toxic signs after infusion was stopped. In contrast, withdrawal of 3,4-DAP infusion at 5 d resulted in re-emergence of toxic sign and death within 12 h, confirming antidotal outcomes require sustained 3,4-DAP treatment for longer than 5 d after intoxication. We exploited this novel survival model of lethal botulism to explore neurophysiological parameters of diaphragm paralysis and recovery. While neurotransmission was nearly eliminated at 5 d, neurotransmission was significantly improved at 21 d in 3,4-DAP-infused survivors, although still depressed compared to naïve rats. 3,4-DAP is the first small molecule to reverse systemic paralysis and promote survival in animal models of botulism, thereby meeting a critical treatment need that is not addressed by post-exposure prophylaxis with conventional antitoxin. These data contribute to a growing body of evidence supporting the use of 3,4-DAP to treat clinical botulism.


Radiation damage to lyophilized mineral solutions during electron probe analysis: quantitative study of chlorine loss as a function of beam current-density and sample mass-thickness.
The quantitative effects of beam current-density and sample mass-thickness on the loss of chlorine which occurs from lyophilized solutes of micro-droplets of mineral salt solutions irradiated in an electron probe analyser were studied. Results are reported for chlorine loss from lyophilized deposits with mass-thickness varying between 5 and 50 mg mm-2 for NaCl salts and 5 and 80 mg mm-2 for KCl salts. Electron accelerating voltage was kept constant at 15 kV. The range of beam current-density (I/S, current/sample surface area) was from 0.1 to 1.5 A mm-2. Samples were irradiated for 1200 s. The results show that under some conditions there is a period of stable chlorine signal before chlorine loss occurs. This is observed between 0.1 and 1 A mm-2, for a period which can last several hundred seconds depending on beam current-density and sample mass-thickness. For each value of I/S, however, no stable chlorine signal can be observed for samples whose mass-thickness exceeds a value negatively correlated with I/S. The curves of decrease of characteristic chlorine X-ray signal (expressed as per cent of count rate in the initial counting interval) versus irradiation time can be fitted by the sum of two exponentials with half lives T1 and T2. In NaCl, T1 and T2 values are highly correlated with I/S but not with mass-thickness. In KCl, T1 is correlated only with mass-thickness and T2 only with I/S. Mixing plasma with mineral solutions prevents chlorine loss.


Prevalence of metabolic syndrome in cirrhotics with gastric antral vascular ectasia.
Gastric antral vascular ectasia (GAVE) is characterized by angliodysplastic lesions that can cause upper gastrointestinal bleeding (UGIB). The mechanism behind GAVE and its association with other diseases remains unknown. We investigated the association of metabolic syndrome in cirrhotic GAVE patients when compared to esophageal variceal hemorrhage (EVH) patients. We performed a retrospective review of 941 consecutive esophagogastroduodenoscopies (EGDs) for UGIB at a medical center between 2017 and 2019. The GAVE group consisted of EGD or biopsy diagnosed cirrhotic GAVE patients, and the EVH group consisted of EVH patients with active bleeding or stigmata of recent hemorrhage on EGD. Baseline variables including co-morbidities and cirrhotic etiology were recorded. Continuous variables were compared using Wilcoxon test and categorical variables were compared using Chi-square or Fisher's exact test. Multiple logistic regression analysis evaluated the association between GAVE and covariates. The final cohort had 96 GAVE and 104 EVH patients. Mean BMI was significantly higher in the GAVE cohort (32.6 vs 27.9, p < 0.0001) in addition to diabetes, hypertension, and hyperlipidemia (53.1% vs 37.5%; 76% vs 47.1%; 38.5% vs 14.4%; respectively, all p < 0.05). Non-alcoholic steatohepatitis (NASH) cirrhosis was more prevalent in GAVE than EVH patients (50% vs 24%, p = 0.0001). Multiple logistics regression revealed female sex, increased BMI, hypertension, and hyperlipidemia all having significantly higher risk of GAVE (all p < 0.05). Our data indicates that when compared to cirrhotics patients with EVH, cirrhotics with GAVE have increased risk of metabolic syndrome. This may play a role in the underlying pathophysiology of GAVE.


Application of a versatile instrument for production of cutaneous anesthesia without needle penetration of the skin.
A self-contained jet injector, the Syrjet Mk II, can be used for needleless injection of anesthetic agents into the skin and subcutaneous tissues. Although designed for dental use, its compact size, simplicity, and versatility make it suited for many medical applications where a relatively small anesthetic field is desired, especially in children. It provides an anesthetic area for the introduction of standard hypodermic needles for more extensive tissue infiltration, or for definitive anesthesia for procedures such as marrow aspiration, biopsies, small lesion excision, and in certain field blocks such as digital or facial sensory. It is significantly free of side effects and complications.


Structure of the Bacterial Cytoskeleton Protein Bactofilin by NMR Chemical Shifts and Sequence Variation.
Bactofilins constitute a recently discovered class of bacterial proteins that form cytoskeletal filaments. They share a highly conserved domain (DUF583) of which the structure remains unknown, in part due to the large size and noncrystalline nature of the filaments. Here, we describe the atomic structure of a bactofilin domain from Caulobacter crescentus. To determine the structure, we developed an approach that combines a biophysical model for proteins with recently obtained solid-state NMR spectroscopy data and amino acid contacts predicted from a detailed analysis of the evolutionary history of bactofilins. Our structure reveals a triangular β-helical (solenoid) conformation with conserved residues forming the tightly packed core and polar residues lining the surface. The repetitive structure explains the presence of internal repeats as well as strongly conserved positions, and is reminiscent of other fibrillar proteins. Our work provides a structural basis for future studies of bactofilin biology and for designing molecules that target them, as well as a starting point for determining the organization of the entire bactofilin filament. Finally, our approach presents new avenues for determining structures that are difficult to obtain by traditional means.


Plasmodium vivax infection alters the peripheral immunoregulatory network of CD4 T follicular cells and B cells.
Regulatory and effector cell responses to Plasmodium vivax, the most common human malaria parasite outside Africa, remain understudied in naturally infected populations. Here, we describe peripheral CD4+ T- and B-cell populations during and shortly after an uncomplicated P. vivax infection in 38 continuously exposed adult Amazonians. Consistent with previous observations, we found an increased frequency in CD4+ CD45RA- CD25+ FoxP3+ T regulatory cells that express the inhibitory molecule CTLA-4 during the acute infection, with a sustained expansion of CD21- CD27- atypical memory cells within the CD19+ B-cell compartment. Both Th1- and Th2-type subsets of CXCR5+ ICOShi PD-1+ circulating T follicular helper (cTfh) cells, which are thought to contribute to antibody production, were induced during P. vivax infection, with a positive correlation between overall cTfh cell frequency and IgG antibody titers to the P. vivax blood-stage antigen MSP119 . We identified significant changes in cell populations that had not been described in human malaria, such as an increased frequency of CTLA-4+ T follicular regulatory cells that antagonize Tfh cells, and a decreased frequency of circulating CD24hi CD27+ B regulatory cells in response to acute infection. In conclusion, we disclose a complex immunoregulatory network that is critical to understand how naturally acquired immunity develops in P. vivax malaria.


Jianghu decoction and its active component polydatin inhibit inflammation and fibrotic lesions in the lungs of ILD mice via the AMPK signaling pathway.
Recently, interstitial lung disease (ILD) morbidity and mortality have been increasing with insidious epidemiological characteristics. Jianghu decoction (JH) is an effective Chinese medicine for ILD. We aimed to reveal the material basis and mechanism of action of JH in the treatment of ILD. In this study, an ILD mouse model was constructed with bleomycin. HE staining, transcriptome analysis, parallel reaction monitoring-mass spectrometry (PRM-MS), UPLC‒MS, and western blotting assays were conducted. HE staining results showed that JH effectively reduced inflammation and fibrosis foci in the lungs of the ILD model. Furthermore, transcriptome analysis revealed that JH regulates a set of biological signaling pathways related to immune inflammation and fibrosis. PRM-MS combined with western blotting was applied to detect inflammation and fibrosis involving proteins in lung tissue. JH effectively reversed the aberrant expression of HMGB1, RAGE, SEPTIN4, ACTA2, and ITGAV proteins in the model group. AMPK was identified as the core upstream regulatory protein for JH-mediated ILD regulation. In addition, UHPLC‒MS technology was applied to determine the active ingredients of JH. A total of 80 components were identified from JH, and polydatin (PD) was identified as the active ingredient that effectively alleviated lung fibrosis and inflammatory injury in ILD mice. To illustrate the molecular regulatory network of JH and PD in alleviating lung fibrosis and inflammatory injury, we also examined inflammation and fibrosis-related molecules downstream of the AMPK pathway with RT‒qPCR and western blotting. The results showed that both JH and its active component PD exert synergistic inhibition on pulmonary fibrosis and inflammation. Specifically, the AMPK/PGC1α/PPARγ signaling pathway was activated, and the AMPK/HMGB1/RAGE signaling pathway was inhibited in ILD lungs responding to JH or PD administration.


Maternally perceived fetal movement patterns: The influence of body mass index.
Maternal reports of decreased fetal movements are associated with adverse pregnancy outcome, but there are conflicting data about perception of fetal movements in women with obesity. To compare perceived fetal movements in women with obesity (body mass index [BMI] ≥30 kg/m2) and women with normal BMI (<25 kg/m2). Data from two separate pregnancy studies were used for this analysis; the Healthy Mums and Babies (HUMBA) trial, which recruited women with obesity and the Multicentre Stillbirth Study (MCSS), which recruited women from a general obstetric population. Fetal movement data were collected using identical interviewer-administered questionnaire in each study. We compared fetal movement strength, frequency and pattern between HUMBA and MCSS women with obesity and MCSS women with normal BMI. Participants were 233 women with obesity and 149 with normal BMI. Mean (SD) gestation at interview was similar between groups (36.9 [2.2] vs 36.6 [0.9], P = 0.06). Perceived fetal movement strength and frequency did not differ between groups. In both women with obesity and normal BMI, a diurnal fetal movement pattern was present, with the majority reporting strong or moderate movements in the evening (88.7% vs 99.3%) and at night-time (92.1% vs 93.1%). Women with obesity, were more likely to report strong fetal movements when hungry (29.1% vs 17.7%, P = 0.001) and quiet fetal movements after eating (47.4% vs 32.0%, P = 0.001). In women with obesity compared to normal BMI, strength and frequency of fetal movements were similar, although patterns were altered in relation to maternal meals.


A two-step fermentation of distillers' grains using Trichoderma viride and Rhodopseudomonas palustris for fish feed.
It is important to provide added value or to make full use of the co-product of grains from ethanol production. In order to convert distillers' grains into a high-quality feed, the Trichoderma viride and Rhodopseudomonas palustris fermentation were combined and investigated in this study. The T. viride fermentation was carried out in an aerobic fermentation installation in favoring of the growth of the fungi and the degradation of the cellulose, and then the fermentation of R. palustris was performed to increase the content of protein with an anaerobic installation. After the two step fermentations, the true protein content of dried distiller' grains increased from 11.4 to 33.6 % (w/w) (the content of crude protein from 14.5 to 39.7 %), the crude fiber content decreased from 21.3 to 7.6 % (w/w), the crude fat content increased from 5.5 to 7.9 % (w/w), the crude ash decreased from 14.6 to 10.2 % (w/w), the total phosphorus content increased from 0.4 to 1.2 % (w/w), and the water content was 11.8 % (w/w). The dried and fermented grains contain the R. palustris viable count of 5.3 × 10¹¹ CFU/g dry matter. The results may support a new application of an active photosynthetic bacteria fish feed in fisheries industry and offer a reference for the further study of lignocellulosic materials as raw materials converting into high-quality feed.


Diagnostic accuracy of a simple qualitative ultrasound assessment for the diagnosis of empty stomach in the adult: A supplementary analysis of a prospective observer-blind randomized crossover study.
It has been reported that qualitative ultrasound assessment performed in the semi-upright position had high sensitivity to detect gastric fluid volume >1.5 mL.kg-1 . Nevertheless, the diagnostic accuracy of qualitative assessment for the diagnosis of empty stomach (fluid volume <0.8 mL.kg-1 ) has not been assessed. We aimed to assess the diagnostic accuracy of simple qualitative ultrasound assessment with and without head-of-bed elevation to 45° for the diagnosis of an empty stomach. We also aimed to determine the diagnostic accuracy of a composite ultrasound scale and clinical algorithm. We performed a supplementary analysis of a prospective observer-blind randomized crossover trial in which adult fasting volunteers attended two distinct sessions, with the head-of-bed angled either at 0° or 45°, in a randomized order. Three tests were performed within each session, each corresponding to a different (either 0, 50, 100, 150 or 200 mL) and randomized volume of water; the same volumes were ingested in both sessions, in a randomized order. Ultrasounds were performed 3 min after water ingestion, blindly to the volume ingested. We included 20 volunteers in whom 120 measurements were analyzed. The sensitivity and specificity of the qualitative assessment in the semirecumbent position were 93% (95% CI: 68-100) and 89% (95% CI: 76-96), respectively. The composite scale and clinical algorithm did not have better diagnostic accuracy than the qualitative assessment performed with head-of-bed elevation. Without head-of-bed elevation, the clinical algorithm had significantly higher specificity (98% [95% CI: 88-100]) than qualitative assessment (67% [95% CI: 51-80]; p < 0.05). These results suggest that qualitative assessment in the semirecumbent position had high diagnostic accuracy for the diagnosis of fluid volume <0.8 mL.kg-1 ; this method can be used in clinical practice for reliable diagnosis of empty stomach.


Inhibition of altered Orai1 channels in Müller cells protects photoreceptors in retinal degeneration.
The expressions of ion channels by Müller glial cells (MGCs) may change in response to various retinal pathophysiological conditions. There remains a gap in our understanding of MGCs' responses to photoreceptor degeneration towards finding therapies. The study explores how an inhibition of store-operated Ca2+ entry (SOCE) and its major component, Orai1 channel, in MGCs protects photoreceptors from degeneration. The study revealed increased Orai1 expression in the MGCs of retinal degeneration 10 (rd10) mice. Enhanced expression of oxidative stress markers was confirmed as a crucial pathological mechanism in rd10 retina. Inducing oxidative stress in rat MGCs resulted in increasing SOCE and Ca2+ release-activated Ca2+ (CRAC) currents. SOCE inhibition by 2-Aminoethoxydiphenyl borate (2-APB) protected photoreceptors in degenerated retinas. Finally, molecular simulations proved the structural and dynamical features of 2-APB to the target structure Orai1. Our results provide new insights into the physiology of MGCs regarding retinal degeneration and shed a light on SOCE and Orai1 as new therapeutic targets.


Histological changes during regression induced by retinoic acid in a transplantable rat chondrosarcoma.
Daily oral treatment with retinoic acid (100 mg/kg bodyweight) induced regression of a transplantable rat chondrosarcoma. In a previous biochemical investigation we have shown that the tissue breakdown is preceded by the loss of proteoglycan. The present study describes the histological changes induced by retinoic acid. A decrease in the intensity of metachromatic staining with toluidine blue was noted already after 1 day and the discoloration was almost complete after 4 days correlating with the loss of proteoglycan. Especially in the perichondrium there was a rapid proliferation of fibroblasts and monocytes. Osteoclast-like cells were missing, but tumor nodules were eroded and split up by penetrating perichondrium. After 4 days of treatment larger necrotic areas were found, initially in the center of tumor nodules only. In other areas the majority of tumorous chondroblasts survived. Tumor nodules appeared partly mesenchyma-like with some fibroblast-like cells suggesting a dedifferentiation of chondroblasts by retinoic acid. we believe that tumor regression induced by retinoic acid involved proteoglycan degradation by chondroblasts themselves and chondroclast-like activity of monocytes and fibroblasts.


Shoulder pain in elite swimmers: primarily due to swim-volume-induced supraspinatus tendinopathy.
Shoulder pain in elite swimmers is common, and its pathogenesis is uncertain. HYPOTHESIS/STUDY DESIGN: The authors used a cross-sectional study design to test Jobe's hypothesis that repetitive forceful swimming leads to shoulder laxity, which in turn leads to impingement pain. Eighty young elite swimmers (13-25 years of age) completed questionnaires on their swimming training, pain and shoulder function. They were given a standardised clinical shoulder examination, and tested for glenohumeral joint laxity using a non-invasive electronic laxometer. 52/80 swimmers also attended for shoulder MRI. 73/80 (91%) swimmers reported shoulder pain. Most (84%) had a positive impingement sign, and 69% of those examined with MRI had supraspinatus tendinopathy. The impingement sign and MRI-determined supraspinatus tendinopathy correlated strongly (r(s)=0.49, p<0.00001). Increased tendon thickness correlated with supraspinatus tendinopathy (r(s)=0.37, p<0.01). Laxity correlated weakly with impingement pain (r(s)=0.23, p<0.05) and was not associated with supraspinatus tendinopathy (r(s)=0.14, p=0.32). The number of hours swum/week (r(s)=0.39, p<0.005) and weekly mileage (r(s)=0.34, p=0.01) both correlated significantly with supraspinatus tendinopathy. Swimming stroke preference did not. These data indicate: (1) supraspinatus tendinopathy is the major cause of shoulder pain in elite swimmers; (2) this tendinopathy is induced by large amounts of swimming training; and (3) shoulder laxity per se has only a minimal association with shoulder impingement in elite swimmers. These findings are consistent with animal and tissue culture findings which support an alternate hypothesis: the intensity and duration of load to tendon fibres and cells cause tendinopathy, impingement and shoulder pain.


Seabuckthorn Wuwei Pulvis attenuates chronic obstructive pulmonary disease in rat through gut microbiota-short chain fatty acids axis.
Seabuckthorn Wuwei Pulvis (SWP) is a traditional Mongolian medicine used in China. It is composed of Hippophae rhamnoides (berries, 30 g), Aucklandiae costus Falc. (dry root, 25 g), Vitis vinifera F. Cordifolia (berries, 20 g), Glycyrrhiza uralensis Fisch. (dry root, 15 g), and Gardenia jasminoides J. Ellis (desiccative ripe fruit, 10 g). It is clinically applied in the treatment of chronic cough, shortness of breath and phlegm, and chest distress. Past studies demonstrated that Seabuckthorn Wuwei Pulvis improved lung inflammation and chronic bronchitis in mice. However, the effect of Seabuckthorn Wuwei Pulvis on chronic obstructive pulmonary disease (COPD) in rats and the underlying action mechanism is not fully understood. To evaluate the anti-COPD effect of Seabuckthorn Wuwei Pulvis and investigate whether its ameliorative effect is correlated with the composition of gut microbiota and its metabolites. The effects of Seabuckthorn Wuwei Pulvis on a COPD rat model were established by exposure to lipopolysaccharide (LPS) and smoking. These effects were then evaluated by monitoring the animal weight, pulmonary function, lung histological alteration, and the levels of inflammatory factors (tumor necrotic factor [TNF]-α, interleukin [IL]-8, IL-6, and IL-17). Furthermore, the serum LPS and fluorescein isothiocyanate-dextran levels were detected by using an enzyme-linked immunosorbent assay and fluorescence microplate reader, respectively. Tight junction proteins (ZO-1 and occludin-1) in the small intestine were detected by performing real-time quantitative polymerase chain reactions and Western blotting to evaluate the intestinal barrier function. The contents of short-chain fatty acids (SCFAs) in the feces of rats were determined by gas chromatography-mass spectrometry. 16S rDNA high throughput sequencing was used to investigate the effect of SWP on the gut microbiota of COPD rats. Treatment with low and median doses of SWP significantly increased the pulmonary function (forced expiratory volume [FEV] 0.3, forced vital capacity [FVC], and FEV0.3/FVC), decreased the levels of TNF-α, IL-8, IL-6, and IL-17 in the lung, and attenuated the infiltration of inflammatory cells into the lung. The low and median doses of SWP shaped the composition of gut microbiota, which increased the abundances of Ruminococcaceae, Christensenellaceae, and Aerococcaceae, increased the productions of acetic acid, propionic acid, and butyric acid, and upregulated the expression of ZO-1 and occludin-1 in the small intestine of COPD rats. SWP improved pulmonary functions and inhibited the inflammatory response by shaping the gut microbiota, increasing SCFA production, and strengthening the intestinal barrier function in rats with COPD induced by LPS and smoking.


Immune checkpoint blockade in the treatment of advanced non-small cell lung cancer - predictors of response and impact of previous radiotherapy.
The implementation of immune checkpoint inhibitors (ICI) into the standard care of advanced non-small cell lung cancer (NSCLC) has improved prognosis for this group of patients. However, long-term survival is rare. The aim of the study was to identify predictors of response and, especially, to investigate the impact radiotherapy might have on duration of response. The association between pretreatment patient/tumor characteristics and progression-free survival (PFS), overall survival (OS), and lung cancer-specific survival was investigated in 78 patients receiving an ICI as ≥2nd line treatment for advanced NSCLC, using Cox regression analysis. Due to competing risk, cause-specific deaths were also examined with cumulative incidence plots. Median OS was 12.6 months (95% CI 7.8-18.2) and median PFS 4.1 months (95% CI 3.0-6.2), after median follow-up time of 49.7 months (range 20.9-51.5). Increasing CRP and neutrophil/lymphocyte ratio (NLR), were associated with poor PFS (CRP: HR 1.49, 95% CI 1.12-1.98; NLR: HR 1.59, 95% CI 1.22-1.85) and OS (CRP: HR 1.94, 95% CI 1.47-2.56; NLR: HR 1.54, 95% CI 1.27-1.87). Radiotherapy prior to immunotherapy was not significantly associated with patient outcome. However, when the dataset was split at 6 months of follow-up, to be able to identify early and late predictors of prognosis, we found that patients receiving radiotherapy <6 months prior to immunotherapy had better PFS (HR: 0.27, 95% CI 0.09-0.84) and lung cancer-specific survival (HR: 0.41, 95% CI 0.18-0.95) after the first 6 months of follow-up, while increasing CRP (PFS: HR1.61, 95% CI 1.21-2.14; OS: HR2.04, 95% CI 1.51-2.74) and NLR (PFS: HR 1.57, 95% CI 1.29-1.91; OS: HR 1.63, 95% CI 1.35-1.97) were predictors of poor short-term prognosis. Radiotherapy may be of importance to achieve a long-lasting response to immunotherapy, while indicators of systemic inflammation can help in identifying patients with poor short-term prognosis.


Experimental evaluation of Candonocypris novaezelandiae (Crustacea: Ostracoda) in the biocontrol of Schistosomiasis mansoni transmission.
To test Candonocypris novaezelandiae (Baird) (C. novaezelandiae), sub-class Ostracoda, obtained from the Nile, Egypt for its predatory activity on snail, Biomphalaria alexandrina (B. alexandrina), intermediate host of Schistosoma mansoni (S. mansoni) and on the free-living larval stages of this parasite (miracidia and cercariae). The predatory activity of C. novaezelandiae was determined on B. alexandrina snail (several densities of eggs, newly hatched and juveniles). This activity was also determined on S. mansoni miracidia and cercariae using different volumes of water and different numbers of larvae. C. novaezelandiae was also tested for its effect on infection of snails and on the cercarial production. C. novaezelandiae was found to feed on the eggs, newly hatched and juvenile snails, but with significant reduction in the consumption in the presence of other diet like the blue green algae (Nostoc muscorum). This ostracod also showed considerable predatory activity on the free-living larval stages of S. mansoni which was affected by certain environmental factors such as volume of water, density of C. novaezelandiae and number of larvae of the parasite. The presence of this ostracod in the aquatic habitat led to significant reduction of snail population, infection rate of snails with schistosme miracidia as well as of cercarial production from the infected snails. This may suggest that introducing C. novaezelandiae into the habitat at schistosome risky sites could suppress the transmission of the disease.


The ABC of vitamin D: a qualitative study of the knowledge and attitudes regarding vitamin D deficiency amongst selected population groups.
In Australia, vitamin D supply in food is limited, and sun exposure is the main source of vitamin D. However skin cancer risk is high, and the need to gain some sun exposure for adequate vitamin D is challenging public health messages to use protection in the sun. The complex vitamin D public health message may be confusing the public and, in particular, those at highest risk for vitamin D deficiency. This study explored vitamin D and sun exposure attitudes, knowledge and practices of some groups considered at risk of vitamin D deficiency and those delivering healthy sun exposure messages to children. 52 adults participated in six focus groups. Results corroborated with previous research showing low levels of vitamin D knowledge. Individual and environmental barriers to receiving adequate sun exposure were also identified. The message advocating balanced sun exposure to produce adequate vitamin D needs to be made clearer and be more effectively communicated. Findings provide insights to aid development of appropriate public health messages for safe sun exposure and vitamin D, especially for vulnerable groups.