The management of epithelial ingrowth after laser in situ keratomileusis.
. To investigate the treatment of corneal epithelial ingrowth with flap melt after LASIK. Thirteen eyes of 13 patients with corneal epithelial ingrowth and flap melt after LASIK were treated based on the thickness and the extent of melting of the flap. Nine eyes were treated by lifting the flap and scraping the epithelium in the interface. Three eyes underwent allogenic keratomileusis. The flap was removed in one eye. Recurrence occurred in 2 eyes treated by lifting the flap. The flaps, however, healed normally after retreatment. The grafts healed normally in the allogenic keratomileusis group, except for one eye in which the flap was lost. The corneal epithelium healed over the stromal bed after the flap was removed. Corneal haze of grade 2 was present at 6 months. The uncorrected visual acuity was between 0.3 and 1.0, and the best-corrected visual acuity lost 1 to 3 lines. Corneal epithelial ingrowth with flap melt is a severe complication after LASIK. However, with proper treatment, vision can be salvaged in most cases.


Compression of the deep branch of the ulnar nerve at the adductor hiatus producing pain without muscle atrophy.
Compression of the deep branch of the ulnar nerve distal to the pisohamate hiatus is rare. Two cases of compression of the deep branch at the adductor hiatus are presented. These cases are unique because of pain in the hand without muscle atrophy. One patient has had surgical treatment, and the other patient's symptoms are not severe enough to warrant surgical treatment.


Quantification of DNA Modifications Using Two-Dimensional Ultraperformance Liquid Chromatography Tandem Mass Spectrometry (2D-UPLC-MS/MS).
Our hereby presented methodology is suitable for reliable assessment of the most common DNA modifications which arise as a product of fundamental metabolic processes. 8-oxoguanine, one of the oxidatively modified DNA bases is a typical biomarker of oxidative stress. A noncanonical base, uracil, may also be present in small quantities in DNA. Ten-eleven translocation (TET) proteins are involved in oxidation of 5-methylcytosine to 5-hydroxymethylcytosine which can be further oxidized to 5-formylcytosine and 5-carboxycytosine. 5-hydroxymethyluracil may be formed in deamination reaction of 5-hydroxymethylcytosine or can also be generated by TET enzymes. All the above mentioned modifications seem to play some regulatory roles. Here, we provide a protocol for isotope-dilution automated online two-dimensional ultraperformance liquid chromatography with tandem mass spectrometry (2D-UPLC-MS/MS) for direct measurement of 5-methyl-2'-deoxycytidine, 5-(hydroxymethyl)-2'-deoxycytidine, 5-formyl-2'-deoxycytidine, 5-carboxy-2'-deoxycytidine, 5-(hydroxymethyl)-2'-deoxyuridine, 2'-deoxyuridine, and 8-oxo-2'-deoxyguanosine. We also provide optimized protocols for extraction of DNA, fully compatible with the downstream MS/MS analysis.


Growth characteristics of clonal cell populations constituting a B16 melanoma metastasis model system.
Three distinct dissemination-related phenotypes have been distinguished among cell subpopulations of the mouse B16 melanoma: tumorigenicity, spontaneous metastasis from subcutaneous tumors, and organ colonization following intravenous injection of cells. From a progenitor clone (G3) of tumorigenic but nonmetastatic and noncolonizing (null) cells that underwent phenotypic diversification in vitro and in vivo, 4 subclones were obtained: G3.5 (culture-generated metastatic), G3.12 (tumor-generated metastatic), G3.15 (culture-generated null), and G3.26 (tumor-generated colonizing). The growth potentials of the parent clone and derived subclones were investigated comparatively in in vivo assays (tumorigenicity, tumor growth rate, and lung colonization potential), monolayer culture assays (generation time, saturation density, clonogenicity, and rate of detachment by trypsin), and in soft agar. In overall growth potential, G3.26 greater than G3.12 greater than G3, G3.5 greater than G3.15. These results indicate that metastatic populations of the B16 melanoma are not the most rapidly and effectively growing cells obtainable from that tumor.


Selenoprotein K Mediates the Proliferation, Migration, and Invasion of Human Choriocarcinoma Cells by Negatively Regulating Human Chorionic Gonadotropin Expression via ERK, p38 MAPK, and Akt Signaling Pathway.
Selenoprotein K (SelK), a member of selenoprotein family, is identified as a single endoplasmic reticulum (ER) transmembrane protein. Although over-expression of SelK inhibits adherence and migration of human gastric cancer BGC-823 cells, the effects of SelK in human choriocarcinoma (CCA) are not well understood. In this study, the expression levels of SelK in three CCA cell lines, BeWo, JEG-3, and JAR, were examined. The effects of silencing or over-expressing SelK on expression of human chorionic gonadotropin beta subunit (β-hCG) were detected by western blotting. The results show that the protein level of β-hCG was reciprocally regulated by down- or up-regulation of SelK (*P < 0.05; #P < 0.05). The proliferative, migratory, and invasive capabilities of JEG-3 cells with reduced or over-expressed SelK were then tested using the cell counting kit-8 (CCK-8), wound healing, and transwell chamber assays. We found that these cellular activities were markedly increased by the loss of SelK in JEG-3 cells. Conversely, over-expressing SelK in JEG-3 cells suppressed these phenotypes. In addition, SelK expression after down- or up-regulation of β-hCG was also measured. Surprisingly, we found that level of SelK was affected by β-hCG (*P < 0.05; #P < 0.05). The proliferation, migration, and invasion were determined in JEG-3 cells after each over-expression and reduction of β-hCG. The results confirmed that β-hCG functions as a promoter of human choriocarcinoma. Furthermore, ERK/p38 MAPK and Akt signaling pathways were found to involve in these cellular functions. This work suggests that SelK may act as a tumor suppressor in human choriocarcinoma cells by negatively regulating β-hCG expression via ERK, p38 MAPK, and Akt signaling pathways. These findings revealed that selenoprotein K may serve as a novel target for human choriocarcinoma therapy in vitro.


Continuous noninvasive measurement of aortic blood flow in critically ill patients with a new esophageal echo-Doppler system.
Determination of aortic blood flow (ABF) using esophageal Doppler has been proposed as a low invasive hemodynamic monitoring method. The esophageal echo-Doppler Dynemo 3000 (Sometec Inc., Paris, France) system, recently available on the market, is an original device measuring simultaneously, and at the same anatomic level, aortic diameter, and blood flow velocity. Until now, this material has been used exclusively for peroperative monitoring. The objectives of the study were to assess the feasibility and reliability of use for continuous measurements of ABF in hemodynamically compromised intensive care unit patients; and to compare ABF values and its change induced by preload manipulation with the cardiac output (CO) values measured simultaneously by the standard thermodilution method. Sixty simultaneous measurements of ABF and CO were performed in 22 intensive care unit patients. In 16 hypovolemic patients, Doppler and thermodilution measurements were repeated after fluid replacement. Applicability of the method was 84.6% (failure of the echo-Doppler method in 4 of 26 eligible patients). Coefficient of variation of echo-Doppler-derived ABF was 3.25 +/- 2.26%. Interobserver variability was 3.3 +/- 1.6%. Close linear relationship was found between ABF and CO (r = 0.92). Average ABF/CO ratio was 73 +/- 10%, but significant variation was observed after fluid replacement. The echo-Doppler Dynemo 3000 system allows reliable continuous measurements of ABF in intensive care unit patients, both easily and safely.


A Review of Super-Resolution Single-Molecule Localization Microscopy Cluster Analysis and Quantification Methods.
Single-molecule localization microscopy (SMLM) is a relatively new imaging modality, winning the 2014 Nobel Prize in Chemistry, and considered as one of the key super-resolution techniques. SMLM resolution goes beyond the diffraction limit of light microscopy and achieves resolution on the order of 10-20 nm. SMLM thus enables imaging single molecules and study of the low-level molecular interactions at the subcellular level. In contrast to standard microscopy imaging that produces 2D pixel or 3D voxel grid data, SMLM generates big data of 2D or 3D point clouds with millions of localizations and associated uncertainties. This unprecedented breakthrough in imaging helps researchers employ SMLM in many fields within biology and medicine, such as studying cancerous cells and cell-mediated immunity and accelerating drug discovery. However, SMLM data quantification and interpretation methods have yet to keep pace with the rapid advancement of SMLM imaging. Researchers have been actively exploring new computational methods for SMLM data analysis to extract biosignatures of various biological structures and functions. In this survey, we describe the state-of-the-art clustering methods adopted to analyze and quantify SMLM data and examine the capabilities and shortcomings of the surveyed methods. We classify the methods according to (1) the biological application (i.e., the imaged molecules/structures), (2) the data acquisition (such as imaging modality, dimension, resolution, and number of localizations), and (3) the analysis details (2D versus 3D, field of view versus region of interest, use of machine-learning and multi-scale analysis, biosignature extraction, etc.). We observe that the majority of methods that are based on second-order statistics are sensitive to noise and imaging artifacts, have not been applied to 3D data, do not leverage machine-learning formulations, and are not scalable for big-data analysis. Finally, we summarize state-of-the-art methodology, discuss some key open challenges, and identify future opportunities for better modeling and design of an integrated computational pipeline to address the key challenges.


Hereditary pediatric cataract on the Arabian Peninsula.
Hereditary pediatric cataract on the Arabian Peninsula does not follow the same epidemiological patterns as described for Western populations. This article describes selected genetic causes for inherited pediatric cataract in the region.


Brain-specific TRAF7 deletion ameliorates traumatic brain injury by suppressing MEKK3-regulated glial inflammation and neuronal death.
Neuronal death and neuroinflammation play critical roles in regulating the progression of traumatic brain injury (TBI). However, associated pathogenesis has not been fully understood. Tumor necrosis factor receptor-associated factor 7 (TRAF7), as the unique noncanonical member of the TRAF family, mediates various essential biological processes. Nevertheless, the effects of TRAF7 on TBI are still unclear. In this study, we showed that TRAF7 expression was markedly up-regulated in cortex and hippocampus of mice after TBI. Brain-specific TRAF7 deletion markedly ameliorated neuronal death in cortical and hippocampal samples of TBI mice, accompanied with cognitive impairments and motor dysfunction. Moreover, the aberrant activation of astrocyte and microglia in cortex and hippocampus of TBI mice was significantly restrained by TRAF7 conditional knockout in brain, as indicated by the increased expression of GFAP and Iba1. In addition, the releases of pro-inflammatory factors caused by TBI were also considerably diminished by brain-specific TRAF7 knockout, which were largely through the blockage of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) signaling pathways. Importantly, mitogen-activated protein kinase kinase kinase 3 (MEKK3) expression levels were greatly enhanced in cortex and hippocampus of mice with TBI, while being dramatically ameliorated by TRAF7 knockout in brain. Mechanistically, we showed that TRAF7 directly interacted with MEKK3. Of note, MEKK3 over-expression almost abrogated the capacity of TRAF7 knockout to mitigate neuronal death and neuroinflammation in the isolated primary cortical neurons and glial cells upon oxygen-glucose-deprivation/reperfusion (OGD/R) stimulation. Collectively, TRAF7 may be an important molecular switch that leads to TBI in a MEKK3-dependent manner, and can be served as a therapeutic target for TBI treatment.


Recognising symptoms of congenital myasthenic syndromes in children: A guide for paediatricians.
Congenital myasthenic syndromes (CMS) are a rare and diverse group of treatable neuromuscular transmission disorders. Diagnosis is often substantially delayed. This study aimed to identify common symptoms of CMS in children and their manifestation to aid diagnosis and early intervention. We performed a retrospective cohort study, including 18 children (median age 13 years, range 9 years 5 months-18 years 0 month) with CMS. Data on CMS symptoms and their manifestation were extracted from patients' charts and supplemented with parental telephone interviews. Descriptive analyses were used to identify common symptoms. A median diagnostic delay of 4 years and 7 months (interquartile range: 51 months) was observed. Proximal muscle weakness (100%), ptosis (89%), clumsy gait (82%), difficulty eating solid foods (78%) and recurrent respiratory tract infections (72%) were most common in these patients. Symptoms mostly co-occurred and frequently had a fluctuating character, aggravated by infections or fatigue. Early referral to diagnose CMS is crucial to enable timely initiation of treatment. Heightened attention to a combination of symptoms related to muscle weakness, rather than individual symptoms, should support paediatricians in flagging these neuromuscular disorders. Medical history taking should be tailored to parents' perceptions, asking questions about recognisable symptoms of muscle weakness.


Exploratory Prognostic Biomarkers of Complement-Mediated Thrombotic Microangiopathy (CM-TMA) in Adults with Atypical Hemolytic Uremic Syndrome (aHUS): Analysis of a Phase III Study of Ravulizumab.
Clinically validated biomarkers for monitoring of patients with complement-mediated thrombotic microangiopathy (CM-TMA) including atypical hemolytic uremic syndrome (aHUS) are unavailable. Improved characterization of biomarkers in patients with aHUS may inform treatment and monitoring for patients with CM-TMA. This analysis used data collected from 55/56 (98.2 %) adult patients with aHUS enrolled in the global Phase III study of ravulizumab (NCT02949128). Baseline (pre-treatment) patient serum, plasma and urine biomarker levels were compared with the maximum observed levels in normal donors and evaluated for associations with pre-treatment plasma exchange/infusion and dialysis status. Biomarkers were also assessed for associations with key clinical measures at baseline and with changes at 26 and 52 weeks from treatment initiation via linear regression analyses. Complement-specific urine levels (factor Ba and sC5b-9) were elevated in >85 % of patients and are significantly associated with pre-treatment kidney dysfunction. Baseline levels of other evaluated biomarkers were elevated in >70 % of patients with aHUS, except for plasma sC5b-9 and serum sVCAM-1. Lower levels of urine complement markers at baseline are significantly associated with improvements in total urine protein and estimated glomerular filtration rate at 26 and 52 weeks of treatment. Clinical assessment of complement activation by a receiver operating characteristic analysis of Ba and sC5b-9 was more sensitive and specific in urine matrix than plasma. This analysis identified a set of biomarkers that may show utility in the prognosis of CM-TMA, including their potential for measuring and predicting response to anti-C5 therapy. Further studies are required to enhance patient risk stratification and improve management of these vulnerable patients. NCT02949128, ClinicalTrials.gov.


Different CXCR4 expression according to various histologic subtype of papillary thyroid carcinoma.
Functional chemokine (C-X-C motif) receptor 4 (CXCR4) is well known to be over-expressed in papillary thyroid carcinoma (PTC). The aim of this study was to evaluate whether or not the expression of CXCR4 is different by histological subtypes of PTC and to elucidate the relationship between the expression of CXCR4 and clinicopathologic factors. CXCR4 expression in 127 PTC samples was assessed using immunohistochemical staining. The expression of CXCR4 showed different patterns according to the histological subtype of PTC (p < 0.001). A strong expression of CXCR4 was observed more frequently in the poorly differentiated region of PTC (81.0 %) than in classical PTC (50.0 %). Strong CXCR4 expression was less frequently shown in follicular variant (33.9 %) and in diffuse sclerosing variant (14.3 %) of PTC. CXCR4 expression showed a distinct pattern according to the histological subtype of PTC although not associated with other clinicopathological parameters.


Circadian rhythm of cat flea (Siphonaptera: Pulicidae) locomotion unaffected by ultrasound.
Cat fleas, Ctenocephalides felis (Bouché), had a circadian rhythm of locomotion that peaked during the last 2 h of the photophase and declined to a lower level that was maintained throughout the scotophase. Activity was lowest during the first 8 h of photophase. The circadian rhythm of these fleas was not affected by an ultrasonic pest control device.


Seven day administration of lamotrigine in epilepsy: placebo-controlled add-on trial.
A double-blind placebo-controlled trial of 7 days administration of lamotrigine as add-on therapy was performed in 10 patients with frequent therapy-resistant, chiefly partial, seizures. Dosage was adjusted on the basis of estimated half-life. Six patients showed a 50% seizure reduction on lamotrigine and two an increase. Side effects (ataxia, dizziness and apathy) occurred in 3 patients, but only at blood levels above 3 micrograms/ml, and were rapidly relieved when the dose was reduced in two. EEG spike counts were significantly reduced on lamotrigine. There was no evidence of interactions with co-medication.


Visualization of experimental glioma C6 by MRI with magnetic nanoparticles conjugated with monoclonal antibodies to vascular endothelial growth factor.
We developed a method for obtaining iron oxide nanoparticles and their conjugation with monoclonal antibodies to vascular endothelial growth factor. The resultant vector nanoparticles were low-toxic and the antibodies retained their immunochemical activity after conjugation. The study was carried out on rats with intracranial glioma C6 on day 14 after its implantation. The intravenously injected nanoparticles visualized the brain tumor in contrast to nanoparticles conjugated with nonspecific immunoglobulins that did not accumulate in the tumor.


Heterologous expression of a fungal sterol esterase/lipase in different hosts: Effect on solubility, glycosylation and production.
Ophiostoma piceae secretes a versatile sterol-esterase (OPE) that shows high efficiency in both hydrolysis and synthesis of triglycerides and sterol esters. This enzyme produces aggregates in aqueous solutions, but the recombinant protein, expressed in Komagataella (synonym Pichia) pastoris, showed higher catalytic efficiency because of its higher solubility. This fact owes to a modification in the N-terminal sequence of the protein expressed in Pichia pastoris, which incorporated 4-8 additional amino acids, affecting its aggregation behavior. In this study we present a newly engineered P. pastoris strain with improved protein production. We also produced the recombinant protein in the yeast Saccharomyces cerevisiae and in the prokaryotic host Escherichia coli, corroborating that the presence of these N-terminal extra amino acids affected the protein's solubility. The OPE produced in the new P. pastoris strain presented the same physicochemical properties than the old one. An inactive form of the enzyme was produced by the bacterium, but the recombinant esterase from both yeasts was active even after its enzymatic deglycosylation, suggesting that the presence of N-linked carbohydrates in the mature protein is not essential for enzyme activity. Although the yield in S. cerevisiae was lower than that obtained in P. pastoris, this work demonstrates the importance of the choice of the heterologous host for successful production of soluble and active recombinant protein. In addition, S. cerevisiae constitutes a good engineering platform for improving the properties of this biocatalyst.


A systematic approach for introduction of novel treatments to a chronic patient group: sacubitril-valsartan as a case study.
To develop a model for systematic introduction and to test the feasibility in a chronic disease population. We also investigated how the approach was received by the patients. The systematic introduction approach is a seven-step procedure: step 1, define a few main criteria; step 2, primary scan patients with the one or two main criteria using computerized medical records/databases/clinical registries; step 3, identify patients applying the other predefined criteria; step 4, evaluate if any examinations/laboratory test updates are required; step 5, summon identified patients to the clinic with an information letter; step 6, discuss treatment with the patient and prescribe if appropriate; and step 7, follow up on initiated therapy and evaluate the applied process. The model was tested in a case study during introduction of the new drug sacubitril-valsartan in a heart failure population. In total, 76 out of 1924 patients were identified to be eligible for sacubitril-valsartan and summoned to the clinic to discuss treatment. Patient experiences with the approach were investigated in an interview study with general inductive approach using qualitative content analysis. This resulted in three final categories: a good approach, role of the information letter, and trust in care. The systematic introduction approach ensures that strict criteria are used in the selection process and that a treatment can be implemented in eligible patients within a specified population with limited resources and time. The model was effective in our case study and maintained the patient's confidence in healthcare.


Attractivity of male rats induced by estradiol and progesterone.
From the present study it appears that castrated male rats can be made attractive to intact males by estradiol benzoate plus progesterone (EB + P). For the determination of attractivity a residential plus-maze was used in which the resident male could choose between the company of the experimental male and that of an ovariectomized female with or without EB + P. In one experiment the males were castrated in adulthood; these animals were less attractive than the ovariectomized females with a similar hormone treatment. In a second experiment it was found that males that had been castrated at birth were as attractive as the ovariectomized females, after treatment with EB + P. These findings indicate that the attractivity that can be induced by EP + P in gonadectomized adult rats is somewhat but not wholly reduced if androgen is present during the neonatal period.


Evaluating the antioxidative defense response of selected indoor plants against benzene and formaldehyde.
Volatile organic compounds (VOCs) such as formaldehyde and benzene are among the key contributors to indoor air pollution. The current situation of environmental pollution is alarming, especially indoor air pollution is becoming a challenge as affecting plants and humans. VOCs are known to adversely affect indoor plants by causing necrosis and chlorosis. In order to withstand these organic pollutants, plants are naturally equipped with an antioxidative defense system. The current research study aimed to evaluate the combined effect of formaldehyde and benzene on the antioxidative response of selected indoor C3 plants including Chlorophytum comosum, Dracaena mysore, and Ficus longifolia. After the combined application of different levels (0, 0; 2, 2; 2, 4; 4, 2; and 4, 4 ppm) of benzene and formaldehyde respectively, in an airtight glass chamber, the enzymatic and non-enzymatic antioxidants were analyzed. Analysis of total phenolics showed a significant increase (10.72 mg GAE/g) in F. longifolia; C. comosum (9.20 mg GAE/g); and D. mysore (8.74 mg GAE/g) compared to their respective controls as 3.76, 5.39, and 6.07 mg GAE/g. Total flavonoids (724 µg/g) were reported in control plants of F. longifolia which were increased to 1545.72 µg/g from 724 µg/g (in control) followed by 322.66 µg/g in D. mysore (control having 167.11 µg/g). Total carotenoid content also increased in D. mysore (0.67 mg/g) followed by C. comosum (0.63 mg/g) in response to increasing the combined dose compared to their control plants having 0.62 and 0.24 mg/g content. The highest proline content was exhibited by D. mysore (3.66 μg/g) as compared to its respective control plant (1.54 μg/g) under a 4 ppm dose of benzene and formaldehyde. A significant increase in enzymatic antioxidants including total antioxidants (87.89%), catalase (59.21 U/mg of protein), and guaiacol peroxidase (52.16 U/mg of protein) was observed in the D. mysore plant under a combined dose of benzene (2 ppm) and formaldehyde (4 ppm) with respect to their controls. Although experimental indoor plants have been reported to metabolize indoor pollutants, the current findings indicate that the combined application of benzene and formaldehyde is also affecting the physiology of indoor plants as well.


[Experimental studies of enzyme activity in the large intestine].
1. A kallikrein is isolated from the human colon. 2. A collagenolytic enzyme is detectable in this tissue. 3. On day 3 after surgery on the colon of the rat, an enhanced collagenolytic activity is demonstrated.


High density lipoproteins down-regulate transcriptional expression of pro-inflammatory factors and oxidative burst in head kidney leukocytes from rainbow trout, Oncorhynchus mykiss.
Teleosts are the first group of vertebrates possessing an acquired immune system; however, it is less developed than in mammals and is highly influenced by environmental changes. Therefore, innate immunity effectors play a more critical role in survival of pathogen-challenged fish. In a previous study we showed that trout high density lipoprotein (HDL), and its major apolipoprotein (ApoA-I) are widely expressed in primary defense barriers and other immune-relevant tissues, displaying important antibacterial activity in vitro. Here we show that trout HDL inhibits both basal and LPS-induced transcript expression of pro-inflammatory cytokines such as TNF-α and IL-1β, and the acute phase protein serum amyloid A (A-SAA), in head kidney leukocytes (HLK) from rainbow trout. In addition, trout HDL was able to block the respiratory burst of PMA-stimulated HKL, at physiological concentrations and in a dose dependent manner. Moreover, this effect was only partially mimicked by supra-physiologic concentrations of the HDL-transported carotenoid, astaxanthin. These results constitute the first data suggesting that in addition to its antimicrobial activity, HDL would have a relevant immunomodulatory role in salmonid fish.


The PurR mutation of Drosophila melanogaster confers resistance to purine and 2,6-diaminopurine by elevating adenosine deaminase activity.
Media supplemented with purine (7H-imidazo[4,5-d]pyrimidine) or the purine analogue 2,6-diaminopurine (DAP) can be employed to select several classes of purine-resistant variants from mutagenized cultures of Drosophila. One class results in elevated resistance to purine and diaminopurine which is correlated with elevated activity of the enzyme adenosine deaminase (adenosine aminohydrolase = EC 18.104.22.168). The first member of this class, Pur R, maps to position 82 +/- in the right arm of the second chromosome. The Pur R mutation causes an elevation of adenosine deaminase (ADA) enzyme activity, apparently by altering a thermolabile, ADA-specific repressor. Pur R may thus encode a negative regulator of adenosine deaminase activity similar to the ADA-binding protein found in mammalian systems.


The glycosphingolipid composition of the placenta of a blood group P fetus delivered by a blood group Pk1 woman and analysis of the anti-globoside antibodies found in maternal serum.
To further define the molecules that may mediate spontaneous abortion due to maternal-fetal blood group incompatibility within the P blood group system, we have examined the fine specificities of maternal antibodies and the glycolipid antigens from the placenta of a P infant born to a Pk1 mother. Maternal antibodies obtained during therapeutic plasmapheresis were analyzed to determine their reactivities with placental glycolipid extracts on thin-layer plates. Second antibodies specific for IgM, IgG, and IgA revealed immunoglobulins of all of these classes strongly reactive with one major placental glycolipid that comigrates with globoside. GC/MS analysis confirmed that the major P-active pentaglycosylceramide of placenta has the same structure as that previously shown for the P antigen of red blood cells: GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc-Cer. Serum antibodies partially purified by affinity chromatography on globoside-octyl-Sepharose specifically recognize glycolipids that contain terminal GalNAc beta 1-3Gal . . . residues and also recognize the same sequence as an internal determinant in some, but not all, glycolipids with extended globoside core regions. Thus, in the blood group P incompatible fetus, the major P antigen present in placenta has the same carbohydrate structure as the P antigen present in fetal and adult erythrocytes and might be a target for the maternal immune system.


Exogenous Jaagsiekte Sheep Retrovirus type 2 (exJSRV2) related to ovine pulmonary adenocarcinoma (OPA) in Romania: prevalence, anatomical forms, pathological description, immunophenotyping and virus identification.
Ovine pulmonary adenocarcinoma (OPA) is a neoplastic disease caused by exogenous Jaagsiekte Sheep Retrovirus (exJSRV). The prevalence of JSRV-related OPA in Eastern European countries, including Romania is unknown. We aimed to investigate: the prevalence and morphological features of OPA (classical and atypical forms) in the Transylvania region (Romania), the immunophenotype of the pulmonary tumors and their relationships with exJSRV infection. A total of 2693 adult ewes slaughtered between 2017 and 2019 in two private slaughterhouses from Transylvania region (Romania) was evaluated. Lung tumors were subsequently assessed by cytology, histology, immunocytochemistry, immunohistochemistry, electron microscopy and DNA testing. Out of 2693 examined sheep, 34 had OPA (1.26% prevalence). The diaphragmatic lobes were the most affected. Grossly, the classical OPA was identified in 88.24% of investigated cases and the atypical OPA in 11.76% that included solitary myxomatous nodules. Histopathology results confirmed the presence of OPA in all suspected cases, which were classified into acinar and papillary types. Myxoid growths (MGs) were diagnosed in 6 classical OPA cases and in 2 cases of atypical form. Lung adenocarcinoma was positive for MCK and TTF-1, and MGs showed immunoreaction for Vimentin, Desmin and SMA; Ki67 expression of classical OPA was higher than atypical OPA and MGs. JSRV-MA was identified by IHC (94.11%) in both epithelial and mesenchymal cells of OPA. Immunocytochemistry and electron microscopy also confirmed the JSRV within the neoplastic cells. ExJSRV was identified by PCR in 97.05% of analyzed samples. Phylogenetic analysis revealed the presence of the exJSRV type 2 (MT809678.1) in Romanian sheep affected by lung cancer and showed a high similarity with the UK strain (AF105220.1). In this study, we confirmed for the first time in Romania the presence of exJSRV in naturally occurring OPA in sheep. Additionally, we described the first report of atypical OPA in Romania, and to the best of our knowledge, in Eastern Europe. Finally, we showed that MGs have a myofibroblastic origin.


Blockade of progesterone-induced increase in hypothalamic luteinizing hormone-releasing hormone levels and serum gonadotropins by intrahypothalamic implantation of 6-hydroxydopamine.
The present study was undertaken to identify the hypothalamic locus where norepinephrine (NE) nerve terminals communicate with luteinizing hormone-releasing hormone (LH-RH)-containing neurons involved in the stimulatory feedback action of gonadal steroids on LH and FSH release. Ovariectomized rats received estradiol benzoate (10 microgram/rat s.c.) on day 0. Intracranial implants containing either 6-hydroxydopamine (6-OHDA), to destroy NE terminals, or cocoa butter (controls) were placed bilaterally in the suprachiasmatic nucleus (SCN), medial basal hypothalamus (MBH) or olfactory tubercle (OT) on day 1. Progesterone (P, 5 mg/rat s.c.) was administered at 10.00 h on day 2 to elicit increases in serum LH and FSH and the MBH LH-RH levels in the afternoon. Implantation of 6-OHDA in the SCN resulted in a marked depletion of NE in and around the region of the SCN in the preoptic-anterior hypothalamic area (POA-AH) without adversely affecting dopamine (DA) concentrations, and blocked the P-induced afternoon increase in the MBH LH-RH and serum gonadotropin levels. Similar reduction in the MBH NE concentrations occurred following placement of 6-OHDA in the MBH; however, these as well as implants in the OT were ineffective in suppressing the P-induced effects. These studies show that functional integrity of the SCN regions is critical in manifestation of the P-induced rise in the MBH LH-RH activity, and this region in the POA-AH, therefore, may be the primary locus of synaptic communication between NE terminals and LH-RH neurons.


Longitudinal detection of subcategorized CD44v6+ CTCs and circulating tumor endothelial cells (CTECs) enables novel clinical stratification and improves prognostic prediction of small cell lung cancer: A prospective, multi-center study.
Current management of small cell lung cancer (SCLC) remains challenging. Effective biomarkers are needed to subdivide patients presenting distinct treatment response and clinical outcomes. An understanding of heterogeneous phenotypes of aneuploid CD31- circulating tumor cells (CTCs) and CD31+ circulating tumor endothelial cells (CTECs) may provide novel insights in the clinical management of SCLC. In the present translational and prospective study, increased cancer metastasis-related cell proliferation and motility, accompanied with up-regulated mesenchymal marker vimentin but down-regulated epithelial marker E-cadherin, were observed in both lentivirus infected SCLC and NSCLC cells overexpressing the stemness marker CD44v6. Aneuploid CTCs and CTECs expressing CD44v6 were longitudinally detected by SE-iFISH in 120 SCLC patients. Positive detection of baseline CD44v6+ CTCs and CD44v6+ CTECs was significantly associated with enhanced hepatic metastasis. Karyotype analysis revealed that chromosome 8 (Chr8) in CD44v6+ CTCs shifted from trisomy 8 towards multiploidy in post-therapeutic patients compared to pre-treatment subjects. Furthermore, the burden of baseline CD44v6+ CTCs (t0) or amid the therapy (t1-2), the ratio of baseline CD31+ CTEC/CD31- CTC (t0), and CTC-WBC clusters (t0) were correlated with treatment response and distant metastases, particularly brain metastasis, in subjects with limited disease (LD-SCLC) but not in those with extensive disease (ED-SCLC). Multivariate survival analysis validated that longitudinally detected CD44v6+/CD31- CTCs was an independent prognostic factor for inferior survival in SCLC patients. Our study provides evidence for the first time that comprehensive analyses of CTCs, CTECs, and their respective CD44v6+ subtypes enable clinical stratification and improve prognostic prediction of SCLC, particularly for potentially curable LD-SCLC.


Structured analyses of interventions to prevent delirium.
Delirium is one of the most serious complications in hospitalized elderly, with incidences ranging from 3-56%. The objective of this meta-analysis was two-fold, first to investigate if interventions to prevent delirium are effective and second to explore which factors increase the effectiveness of these interventions. An electronic search was carried out on articles published between January 1979 and July 2009. Abstracts were reviewed, data were extracted and methodologic quality was assessed by two independent reviewers. Effect sizes of the interventions were expressed as ORs (odds ratios) and 95%CIs (confidence intervals). A random effect model was used to provide pooled ORs. To explore which factors increase the effectiveness of the interventions, ORs were stratified for several factors. Sixteen relevant studies were found. Overall the included studies showed a positive result of any intervention to prevent delirium (pooled OR: 0.64; 95%CI: 0.46-0.88). The largest effect was seen in studies on populations with an incidence of delirium above 30% in the control group (pooled OR: 0.34; 95%CI: 0.16-0.71 versus 0.76; 95%CI: 0.60-0.97). Interventions to prevent delirium are effective. Interventions seem to be more effective when the incidence of delirium in the population under study is above 30%. To maximize the options for a cost-effective strategy of delirium prevention it might be useful to offer an intervention to a selected population.


Preparation and in-vitro/in-vivo evaluation of curcumin nanosuspension with solubility enhancement.
We developed Cur nanosuspension (Cur-NS) with PVPK30 and SDS as stabilizers to improve poor water solubility and short biological half-time of Cur. Physicochemical characterization of Cur-NS was characterized systematically. The in-vitro dissolution, cytotoxicity and in-vivo pharmacokinetic experiments of Cur-NS were also evaluated. Scanning electron microscope indicated that the morphologies of Cur-NS were spherical or ellipsoidal in shape. X-ray diffraction verified that Cur was successfully developed as nanoparticles with an amorphous phase in Cur-NS. Fourier transform infrared spectroscopy suggested there was no degradation about Cur in the Cur-NS. Furthermore, the in-vitro study showed that the cumulative release of the Cur-NS was 82.16 ± 2.62% within 34 h and the cytotoxicity of the Cur-NS against HepG2 cells was much better than raw Cur. Besides, in-vivo pharmacokinetics in rats by intravenous injection displayed that the in-vivo process of Cur-NS pertained to two-compartment model. Meanwhile, the t1/2 and AUC0-t of Cur-NS were enhanced by 11.0-fold and 4.2-fold comparing to Cur solution. The Cur-NS significantly increased the water solubility and half-time of Cur, suggesting its potential as a nanocarrier in the delivery of Cur for future clinical application.


Diminution of signal transducer and activator of transcription 3 signaling inhibits vascular permeability and anaphylaxis.
During IgE-mediated immediate hypersensitivity reactions, vascular endothelial cells permeabilize in response to mast cell mediators. We have demonstrated previously that patients and mice with signal transducer and activator of transcription 3 (STAT3) mutations (autosomal dominant hyper-IgE syndrome [AD-HIES]) are partially protected from anaphylaxis. We sought to study the mechanism by which STAT3 contributes to anaphylaxis and determine whether small-molecule inhibition of STAT3 can prevent anaphylaxis. Using unaffected and STAT3-inhibited or genetic loss-of-function samples, we performed histamine skin prick tests, investigated the contribution of STAT3 to animal models of anaphylaxis, and measured endothelial cell permeability, gene and protein expression, and histamine receptor-mediated signaling. Although mouse mast cell degranulation was minimally affected by STAT3 blockade, mast cell mediator-induced anaphylaxis was blunted in Stat3 mutant mice with AD-HIES and in wild-type mice subjected to small-molecule STAT3 inhibition. Histamine skin prick test responses were diminished in patients with AD-HIES. Human umbilical vein endothelial cells derived from patients with AD-HIES or treated with a STAT3 inhibitor did not signal properly through Src or cause appropriate dissolution of the adherens junctions made up of the proteins vascular endothelial-cadherin and β-catenin. Furthermore, we found that diminished STAT3 target microRNA17-92 expression in human umbilical vein endothelial cells from patients with AD-HIES is associated with increased phosphatase and tensin homolog (PTEN) expression, which inhibits Src, and increased E2F transcription factor 1 expression, which regulates β-catenin cellular dynamics. These data demonstrate that STAT3-dependent transcriptional activity regulates critical components for the architecture and functional dynamics of endothelial junctions, thus permitting vascular permeability.


Membrane cut-off does not influence results regarding the measurement of small molecules - a comparative study between 20- and 100-kDa catheters in hepatic microdialysis.
Microdialysis is a method used to monitor hepatic tissue metabolism. Membranes with a molecular cut-off of 20 kilodalton (kDa) are currently used to measure the small metabolites glucose, glycerol, lactate and pyruvate. Using membranes with higher cut-off such as 100 kDa allows the possibility of measuring larger molecules but may affect results regarding small molecules. The aim was to compare microdialysis catheters with a cut-off of 20 and 100 kDa in the measurement of small molecules in a pig liver model. Four microdialysis catheters were inserted into the liver of each pig used in the experiment (n = 6). Two catheters with cut-off of 20 kDa were perfused with Ringer acetate, and two catheters with cut-off of 100 kDa: one perfused with Ringer acetate and one with hydroxyethyl starch (Voluven) at a flow rate of 0·3 μl min(-1). Dialysate samples were collected at 40-min intervals and analysed for glucose, glycerol, lactate and pyruvate. Compared to the other catheters, the 100-kDa catheters perfused with Ringer acetate tended to measure higher dialysate concentrations of the small molecules, the difference reaching statistical significance in the case of pyruvate. Concentrations measured by the 100-kDa catheters perfused with Voluven were, however, comparable to the 20-kDa catheters. Microdialysis catheters with membrane cut-off of 20 and 100 kDa can be used equally in hepatic microdialysis for the monitoring of glucose, glycerol, lactate and pyruvate, and lactate/pyruvate ratio if a high osmotic solution (Voluven) is used to perfuse the 100-kDa catheters.


Allele-specific PCR with a novel data processing method based on difference value for single nucleotide polymorphism genotyping of ALDH2 gene.
Single nucleotide polymorphism (SNP) analysis based on allele-specific polymerase chain reaction (AS-PCR) is a relatively effective and economical method compared with other genotyping technologies such as DNA sequencing, DNA hybridization and isothermal amplification strategies. But AS-PCR is limited by its labor-intensive optimization of reaction parameters and time-consuming result assessment. In this study, we put forward a novel idea of data processing to address this problem. SNP analysis was accomplished by AS-PCR with endpoint electrochemical detection. For each sample, two separate reactions were run simultaneously with two sets of allele-specific primers (wild-type primers for W system and mutant primers for M system). We measured their redox current signals on screen-printed electrodes once AS-PCR finished and calculated the difference value of current signals between two systems to determine the genotyping result. Based on the difference value of fluorescent signals, real-time fluorescent PCR was used to study reaction parameters in AS-PCR. With screened parameters, we obtained the genotyping results within 50 min. 36 hair-root samples from volunteers were analyzed by our method and their genotypes of ALDH2 gene (encoding aldehyde dehydrogenase 2) were totally identical with data from commercialized sequencing. Our work first employed difference value between two reaction systems to differentiate allele and provided a novel idea of data processing in AS-PCR method. It is able to promote the quick analysis of SNP in the fields of health monitor, disease precaution, and personalized diagnosis and treatment.


Usefulness of Fractal Analysis of Kirsch Edge Images for the Tissue Fragment Inner Structure in Breast FNAB.
Recent advances in high-precision mammography and ultrasound screening have led to an increase in the detection of early lesions (ductal carcinoma in situ and small cancers) appearing as microcalcified lesions or microcystic images, and there needs to be an improvement in the accuracy of breast fine-needle aspiration biopsy (FNAB) assessing these lesions. The objective of this study was to investigate whether fractal analysis of Kirsch edge images for the tissue fragment inner structure (FKT) is useful in breast FNAB. FKT measures tissue fragment chromasia of hyperchromatic crowded tissue fragments (HCG), tissue fragment shape unevenness, and tissue fragment inner structure complexity. Study Design Materials: Nineteen epithelial tissue fragments of fibroadenoma (FA) from 7 patients and 52 tissue fragments of invasive breast carcinoma of no special type (IBC-NST) (grade 1-2) from 11 patients were assessed. First, tissue fragments were classified into small (smaller than 60 × 102 μm2), medium, and large (100 × 102 μm2 or larger), and the appearance rate of each size was determined. Second, for FKT, the luminance value of tissue fragment chromasia, the unevenness and fractal value, and the tissue fragment inner structure complexity were determined. In statistical analysis, the Steel-Dwass test, nonlinear discriminant analysis, and receiver operating characteristic analysis were performed, setting the significance level at p < 0.05. "Unevenness of the tissue fragment shape," "fractal value of the tissue fragment shape," and "fractal value of the tissue fragment inner structure" were significantly higher in small and large tissue fragments in IBC-NST compared with those in FA. The specificity and sensitivity were the highest (100%) in small tissue fragments in multivariate analysis using 4 variables ("luminance value of tissue fragment chromasia," "unevenness of tissue fragment shape," "fractal value of the tissue fragment shape," and "fractal value of the tissue fragment inner structure"). FKT, which evaluates "tissue fragment darkness," "tissue fragment shape unevenness," and "tissue fragment inner structure complexity" focusing on small tissue fragments of HCG in breast FNAB, is useful as a system that assists cytopathological assessment of breast FNAB.


Early reduction in prefrontal theta QEEG cordance value predicts response to venlafaxine treatment in patients with resistant depressive disorder.
Previous studies of patients with unipolar depression have shown that early decrease of prefrontal EEG cordance in theta band can predict clinical response to various antidepressants. We have now examined whether decrease of prefrontal quantitative EEG (QEEG) cordance value after 1 week of venlafaxine treatment predicts clinical response to venlafaxine in resistant patients. We analyzed 25 inpatients who finished 4-week venlafaxine treatment. EEG data were monitored at baseline and after 1 week of treatment. QEEG cordance was computed at three frontal electrodes in theta frequency band. Depressive symptoms and clinical status were assessed using Montgomery-Asberg Depression Rating Scale (MADRS), Beck Depression Inventory-Short Form (BDI-S) and Clinical Global Impression (CGI). Eleven of 12 responders (reduction of MADRS >or=50%) and only 5 of 13 non-responders had decreased prefrontal QEEG cordance value after the first week of treatment (p=0.01). The decrease of prefrontal cordance after week 1 in responders was significant (p=0.03) and there was no significant change in non-responders. Positive and negative predictive values of cordance reduction for response were 0.7 and 0.9, respectively. The reduction of prefrontal theta QEEG cordance value after first week of treatment might be helpful in the prediction of response to venlafaxine.


Ketamine attenuates high mobility group box-1-induced inflammatory responses in endothelial cells.
High mobility group box-1 (HMGB1) acts as an inflammatory mediator and has been implicated in pathophysiological damage of vascular inflammatory diseases. Ketamine, an anesthetic agent with sedative and analgesic properties, has been shown to have potent anti-inflammatory effects in a variety of models of systemic inflammation. However, the effects of ketamine on HMGB1-mediated proinflammatory responses have not been fully investigated. In the present study, we investigated the effects of ketamine on HMGB1-activated endothelial cells and explored the underlying mechanisms. Human endothelial cells were incubated with or without HMGB1 (1 μg/mL) in the presence or absence of ketamine, an nuclear factor (NF)-κB inhibitor (PDTC), anti-toll-like receptor (TLR)2/4 antibody, or small interfering RNA (siRNA). The anti-inflammatory activities of ketamine were determined by measuring solute flux, leukocyte adhesion and migration, and activation of proinflammatory proteins in HMGB1-activated endothelial cells. The effect of ketamine on TLR-2/4 and NF-κB activation was evaluated using enzyme-linked immunosorbent assays and immunofluorescence confocal microscopy assay. We found that ketamine inhibited the HMGB1-mediated barrier disruption, neutrophil adhesion and migration, and expression of cell adhesion molecules in a dose-dependent manner. Furthermore, ketamine downregulated the TLR-2 and -4, expression in HMGB1-activated endothelial cells. Treatment with ketamine also significantly inhibited the activation of TLR2/4 and the nuclear translocation of NF-κB p50/p65. Furthermore, our study shows that the HMGB1-induced release of inflammatory mediators was suppressed by PDTC, anti-TLR2/4 antibody, and siRNA. Our study has demonstrated that ketamine exerts anti-inflammatory effects in HMGB1-mediated proinflammatory responses in a dose-dependent manner. The mechanism responsible for these effects involves the TLR2/4 and NF-κB signaling pathway.


Effect of anterior resection on anal sphincter function.
Minor difficulties with continence may occur after low anterior resection. Intraoperative injury to the internal anal sphincter or its nerve supply may contribute to this. To study the effect of low anterior resection on the anal sphincter mechanism, anal manometry was performed on 20 patients before and 10 days after resection. Fifteen patients were studied again 6 months after operation. Resting, maximum squeeze and squeeze increment pressures were recorded. Intraoperative manometry (n = 11) and presacral nerve stimulation (n = 6) were performed to determine whether peroperative injury to the internal anal sphincter had occurred. Resting and maximum squeeze anal canal pressures were reduced by low anterior resection, and did not recover. The squeeze pressure increment did not change. Division of the inferior mesenteric artery, full mobilization of the rectum and mesorectum, and rectal transection did not affect resting anal pressure, which was reduced after EEA anastomosis (mean (s.e.m.) before, 40(5) mmHg; after, 27(4) mmHg; P less than 0.05, n = 5). Presacral nerve stimulation produced relaxation of the internal sphincter. Anal sphincter pressures are reduced after low anterior resection. The external anal sphincter and the nerve supply to the internal anal sphincter appear intact. A direct injury to the internal sphincter is postulated.


The Case.
KS is a 76-year-old Burmese woman who presented to the hospital with right-sided weakness that had begun two days prior. Although KS had been diagnosed earlier with dementia and had a medical history of stroke, she was living at a nursing home and was able to walk on her own and use her right arm to feed herself. Since her last stroke three years ago, she had also developed aphasia, and her speech has been minimal. During her current hospitalization, KS was found to have a cerebral hemorrhage, but after consultation with neurosurgery, the team determined that no medical interventions were available, and a higher level of care was not required. She also presented with other medical conditions, including severe hypertension, chronic kidney disease, and depression. The patient's blood pressure has remained high. She has now failed a swallow evaluation and is refusing the insertion of a nasogastric feeding tube. The patient has a younger sister who believes, from a cultural perspective, that only the patient should make her medical decisions. As a result, she feels helpless in assisting the team. The medical team has asked for an ethics consultation to direct the patient's care and to guide decisionmaking.


Habitat Diversity, Stability, and Productivity of Malaria Vectors in Irrigated and Nonirrigated Ecosystems in Western Kenya.
Several sub-Saharan African countries rely on irrigation for food production. This study examined the impact of environmental modifications resulting from irrigation on the ecology of aquatic stages of malaria vectors in a semi-arid region of western Kenya. Mosquito larvae were collected from irrigated and non-irrigated ecosystems during seasonal cross-sectional and monthly longitudinal studies to assess habitat availability, stability, and productivity of anophelines in temporary, semipermanent, and permanent habitats during the dry and wet seasons. The duration of habitat stability was also compared between selected habitats. Emergence traps were used to determine the daily production of female adult mosquitoes from different habitat types. Malaria vectors were morphologically identified and sibling species subjected to molecular analysis. Data was statistically compared between the two ecosystems. After aggregating the data, the overall malaria vector productivity for habitats in the two ecosystems was estimated. Immatures of the malaria vector (Anopheles arabiensis) Patton (Diptera: Culicidae) comprised 98.3% of the Anopheles in both the irrigated and non-irrigated habitats. The irrigated ecosystem had the most habitats, higher larval densities, and produced 85.8% of emerged adult females. These results showed that irrigation provided conditions that increased habitat availability, stability, and diversity, consequently increasing the An. arabiensis production and potential risk of malaria transmission throughout the year. The irrigated ecosystems increased the number of habitats suitable for Anopheles breeding by about 3-fold compared to non-irrigated ecosystems. These results suggest that water management in the irrigation systems of western Kenya would serve as an effective method for malaria vector control.


Real-world implementation of precision psychiatry: Transdiagnostic risk calculator for the automatic detection of individuals at-risk of psychosis.
Risk estimation models integrated into Electronic Health Records (EHRs) can deliver innovative approaches in psychiatry, but clinicians' endorsement and their real-world usability are unknown. This study aimed to investigate the real-world feasibility of implementing an individualised, transdiagnostic risk calculator to automatically screen EHRs and detect individuals at-risk for psychosis. Feasibility implementation study encompassing an in-vitro phase (March 2018 to May 2018) and in-vivo phase (May 2018 to April 2019). The in-vitro phase addressed implementation barriers and embedded the risk calculator (predictors: age, gender, ethnicity, index cluster diagnosis, age*gender) into the local EHR. The in-vivo phase investigated the real-world feasibility of screening individuals accessing secondary mental healthcare at the South London and Maudsley NHS Trust. The primary outcome was adherence of clinicians to automatic EHR screening, defined by the proportion of clinicians who responded to alerts from the risk calculator, over those contacted. In-vitro phase: implementation barriers were identified/overcome with clinician and service user engagement, and the calculator was successfully integrated into the local EHR through the CogStack platform. In-vivo phase: 3722 individuals were automatically screened and 115 were detected. Clinician adherence was 74% without outreach and 85% with outreach. One-third of clinicians responded to the first email (37.1%) or phone calls (33.7%). Among those detected, cumulative risk of developing psychosis was 12% at six-month follow-up. This is the first implementation study suggesting that combining precision psychiatry and EHR methods to improve detection of individuals with emerging psychosis is feasible. Future psychiatric implementation research is urgently needed.


Evaluation of dose variation during total skin electron irradiation using thermoluminescent dosimeters.
To determine acceptable dose variation using thermoluminescent dosimeters (TLD) in the treatment of Mycosis Fungoides with total skin electron beam (TSEB) irradiation. From 1983 to 1993, 22 patients were treated with total skin electron beam therapy in the standing position. A six-field technique was used to deliver 2 Gy in two days, treating 4 days per week, to a total dose of 35 to 40 Gy using a degraded 9 MeV electron beam. Thermoluminescent dosimeters were placed on several locations of the body and the results recorded. The variations in these readings were analyzed to determine normal dose variation for various body locations during TSEB. The dose to flat surfaces of the body was essentially the same as the dose to the prescription point. The dose to tangential surfaces was within +/- 10% of the prescription dose, but the readings showed much more variation (up to 24%). Thin areas of the body showed large deviations from the prescription dose along with a large amount of variation in the readings (up to 22%). Special areas of the body, such as the perineum and eyelid, showed large deviations from the prescription dose with very large (up to 40%) variations in the readings. The TLD results of this study will be used as a quality assurance check for all new patients treated with TSEB. The results of the TLDs will be compared with this baseline study to determine if the delivered dose is within acceptable ranges. If the TLD results fall outside the acceptable limits established above, then the patient position can be modified or the technique itself evaluated.


Oxidized LDL induces a coordinated up-regulation of the glutathione and thioredoxin systems in human macrophages.
Using DNA microarray analysis, we found that human macrophages respond to oxidized low-density lipoprotein (oxLDL) by activating the antioxidative glutathione and thioredoxin systems. Several genes of the glutathione and thioredoxin systems were expressed at high levels in macrophages when compared to 80 other human tissues and cell types, indicating that these systems may be of particular importance in macrophages. The up-regulation of three genes in these systems, thioredoxin (P < 0.005), thioredoxin reductase 1 (P < 0.001) and glutathione reductase (P < 0.001) was verified with real-time RT-PCR, using human macrophages from 10 healthy donors. To investigate the possible role of these antioxidative systems in the development of atherosclerosis, expression levels in macrophages from 15 subjects with atherosclerosis (12 men, 3 women) and 15 matched controls (12 men, 3 women) were analyzed using DNA microarrays. Two genes in the glutathione system Mn superoxide dismutase (P < 0.05) and catalase (P < 0.05) differed in expression between the groups. We conclude that macrophage uptake of oxidized LDL induces a coordinated up-regulation of genes of the glutathione and thioredoxin systems, suggesting that these systems may participate in the cellular defense against oxidized LDL and possibly modulate the development of atherosclerosis.


Surgical options in suprastomal collapse-induced severe airway obstruction.
A single institutions experience with various surgical options in the treatment of severe suprastomal collapse (SSC). The study included 18 tracheostomized children with SSC treated between January 2012 and December 2018. Data included: patient demography, initial airway lesions, comorbidities, indication and age at tracheostomy, prior airway surgery, stomal demography, type of surgery, postoperative management, complications and treatment outcomes. Four techniques were used to correct SSC. The surgical choice was dependent on stoma demography and associated airway lesions. Excision was done in eight patients and rib cartilage augmentation in five. Three patients had single stage tracheal resection and anastomosis. Two patients received stomal rigidification and temporary placement of Montgomery T tube. Three patients with anterior rib graft augmentation required additional lateral tracheal wall rigidification. Three patients (two with cartilage augmentation, and one with stomal rigidification) developed minimal granulation tissue in the postoperative period. Complete SSC resolution was seen in all except two patients who had a partial response to the treatment. All patients were successful decannulated and are currently asymptomatic. Decannulation failures may be due to severe suprastomal collapse that could be either unique or associated with obstructing laryngotracheal lesions. Therefore, it is essential to select the most appropriate surgical treatment to obtain overall favorable outcomes.


Binary Pluronics based mixed micellar systems: Effective solution for improved solubilization of Biochanin A.
The present work was carried out to appraise the solubilization of Biochanin (BCA), a natural hydrophobic drug in pure Pluronics (P84, P123 and F127) as well as binary mixed micelles (P84-P123 and F127-P123) followed by an interactional study using various state of art techniques. The critical micelle concentration (CMC) of binary mixtures P84-P123 and F127-P123 was found to be significantly lower than pure Pluronics justifying the candidature of mixed micelles (MMs) as a better solubilizer for BCA. UV-visible studies revealed the enhanced solubility of BCA in P84-P123 (1:2) MMs (3.51 ± 0.087 mg/mL) and F127-P123 (1:2) MMs (2.94 ± 0.046 mg/mL) as compared to micelles of pure Pluronics (10% w/v), P84 (2.25 ± 0.039 mg/mL) and F127 (1.12 ± 0.036 mg/mL). Differential pulse voltammetry (DPV) results demonstrated significantly superior binding of BCA with P84-P123 (1:2) MM (Ka = 2.60 × 105 M-1) as compared to pure P84 (Ka = 1.415 × 105 M-1). Mixed micellization (P84-P123 (1:2)) leads to increase in micellar hydrodynamic diameter (Dh = 16.09 nm) as compared to pure Pluronic P84 micelles (Dh = 15.71 nm) which was further amplified (Dh = 17.69 nm) after BCA loading. Different formulations of pure and MMs were subjected to in vitro drug release and MMs were found to slow down BCA release as compared to pure Pluronics (P84, P123). The results obtained in this study proved that P84-P123 (1:2) MMs are superior and more effective for the solubilization of BCA than pure and other MMs. Thus, the examined MMs hold the potential for advancing the development of solubilization techniques for other hydrophobic drugs with significant pharmacological value.


Host immune responses in hepatitis C virus clearance.
The factors that determine the outcome of hepatitis C virus (HCV) infection are not fully understood. An increased and broadly targeted/multispecific T-cell response is thought to be paramount to a favourable outcome. Human leucocyte antigen (HLA) genes, in particular DRB1 and DQB1, are also reported to influence outcome of infection. We have previously demonstrated strong associations between DRB10101 and spontaneous viral clearance. The aim of the current study was to investigate HCV-specific T-cell response and the influence of DRB10101 in patients with long-term history of HCV clearance as compared to patients that developed persistent HCV infection. The proliferation of peripheral blood mononuclear cells stimulated with five non-structural and core HCV antigens and 20 synthesized HCV peptides, designed using T-cell epitope-predictive software, was determined by the incorporation of H-thymidine. Although HCV-specific T-cell responses were more frequently detected and a broader range of peptides were targeted in the viral clearance group, the magnitude and breadth of the responses were not significantly different to that in the viral persistence group. The magnitude and breadth of the T-cell response was significantly associated, however, with possession of DRB10101. Furthermore DRB10101 positive individuals with viral clearance had broader HCV-specific T-cell responses. These findings lend further credence to the importance of the host immune system to the outcome of HCV infection and provide a rationale for the role of DRB10101 in the resolution of HCV infection.


EGFR exon 21 L858R as an acquired resistance mechanism to nivolumab in a lung cancer patient originally driver gene-negative.
The use of immune checkpoint inhibitors targeting PD-1 and PD-L1 in advanced non-small cell lung cancer (NSCLC) has been one of the most significant improvements in recent years. However the resistance mechanisms of immune checkpoint inhibitors require further investigation. Herein we attempted to determine the possible resistance mechanism of nivolumab in a male smoker with advanced adenosquamous carcinoma. After experiencing disease progression on systematic chemotherapy, he was administered nivolumab as a result of high PD-L1 expression. Larger panel gene detection was performed after the failure of nivolumab treatment to investigate the possible resistance mechanism and a new EGFR exon 21 L858R mutation was detected. After a achieving a response with gefitinib, the patient suffered a rapid relapse and died of tumor progression. This case represents the first time EGFR exon 21 L858R has been detected as an acquired resistance mutation to nivolumab. Patients with high PD-L1 expression may exhibit a poor response to EGFR-tyrosine kinase inhibitors. Large panel gene detection remains the optimal choice when confronted with drug resistance.


AGEs-RAGE axis causes endothelial-to-mesenchymal transition in early calcific aortic valve disease via TGF-β1 and BMPR2 signaling.
Recent studies reported that advanced glycation end products (AGEs) and endothelial-to-mesenchymal transition (EndMT) were involved in the calcific aortic valve disease (CAVD). However, the roles of AGEs in EndMT in the development of CAVD have not been elucidated. In this study, six-week-old male Apoe-/- mice were divided into four groups based on the following feeding periods: 0, 2, 4, and 6 months. The latter three groups were further divided into three subgroups corresponding to the following diet treatments: normal diet, high-fat diet + normal saline injection, and high-fat diet + aminoguanidine injection. Weight gain was monitored weekly. Finally, heart echocardiographic assessments were performed, and serum lipid levels, the protein expression and the histological changes in the aortic valves were determined. Results showed that the AGE inhibitor aminoguanidine alleviated the transaortic valve velocity and decreased the total cholesterol and low-density lipoprotein cholesterol levels. Calcification and carboxymethyl-lysine deposition were firstly detected around the aortic valve surfaces, whereas aminoguanidine injection attenuated their accumulation. In the early stage, HFD-activated AGEs-RAGE signaling resulted in the alpha-smooth muscle actin (α-SMA) upregulation and the vascular endothelium cadherin (VE-cadherin) downregulation on the valve endothelial layer. The activation resulted in early the transforming growth factor-β1 (TGF-β1) and the alkaline phosphatase (ALP) upregulation, and simultaneously reduced the bone morphogenetic protein receptor type II (BMPR2) expression. However, aminoguanidine restricted these proteins changes by inhibiting the interaction of AGEs and RAGE. In addition, immunofluorescence images showed obvious double-positive staining of ALP and α-SMA on the valve surfaces, revealing the contribution of EndMT to the early calcification. Therefore, this study demonstrates that activation of the AGEs-RAGE axis induced EndMT, promoting the progression of the aortic valve calcification in the initial stage via the counteraction of BMPR2 and TGF-β1 signaling.


Blood vessels of the peri-implant mucosa: a comparison between flap and flapless procedures.
The vascularity of a peri-implant tissue is a very important parameter in the establishment and maintenance of healthy tissue after dental implant insertion. The purpose of this study was to compare the vascularity of the peri-implant mucosa between flap and flapless implant surgeries by using a canine mandible model. In six mongrel dogs, bilateral, edentulated, and flat alveolar ridges were created in the mandible. After 3 months of healing, 2 implants were placed in each side by either flap or flapless procedure. After another healing period of 3 months, biopsies were obtained, prepared for light microscopy, and exposed to morphometric measurements. The supracrestal connective tissue lateral to the implant was found to be more richly vascularized in the flapless group than in the flap group. These results suggest that the flapless procedure may increase the vascularity of the peri-implant mucosa.


[Are small-size depressed-type colorectal neoplasias really the expression of a early stage of colorectal cancer. Apropos of a clinical case].
A route of colorectal cancer development other than the adenoma-carcinoma sequence has recently become an issue due to the discovery of the depressed-type early colorectal cancers. Despite the protruding shapes of depressed-type early colorectal cancers, they probably have biological characteristics which differ from those of the usual polyp lesions. They show more aggressive behavior than the polypoid type and can arise de novo. Depressed-type lesions, in contrast to flat-type or protruded-type lesions, tend to invade the submucosa rapidly, so it is better treat them surgically from the outset. We report a case of a small depressed-type colorectal cancer involving the caecum of a 79-year-old male patient.


Benthic foraminifera as bio-indicator of marine pollution in the southwestern Bay of Bengal, India.
The benthic foraminiferal community is an excellent bio-indicator of pollution in coastal and marginal marine settings. Their abundance, diversity and its relationship with environmental parameters, viz. grain size, organic carbon (Corg) and trace metals concentration, from the surface sediment layer of the southwestern shelf of the Bay of Bengal, have been used to understand the marine pollution in this study. A total of 32 surface sediment samples were collected at various water depths.The samples were analysed for grain size distribution, organic matter (OM), foraminiferal and trace metals studies. Statistical analyses such as correlation matrix, PCA (Principal Component Analysis) and cluster analysis show that the foraminiferal species composition is significantly influenced by the organic carbon (Corg), Co, Pb, Zn, Cr, and Ni concentrations. The dominance of stress-tolerant species, viz. Ammonia beccarii, Ammonia tepida, Nonion faba, Bulimina marginata, Bolivina robusta, Elphidium craticulatum and Elphidium advenum, as well as less species diversity, demonstrates the elevated Corg and trace metal pollution in the marine environment.


Oxygen-Ozone Therapy Associated with Alpha Lipoic Acid Plus Palmitoylethanolamide and Myrrh versus Ozone Therapy in the Combined Treatment of Sciatic Pain Due to Herniated Discs: Observational Study on 318 Patients.
The aim of our observational study is to compare the therapeutic efficacy of combined treatment of oxygen-ozone therapy and oral treatment with alpha-lipoic acid (ALA) + palmitoylethanolamide (PEA) and myrrh in patients with peripheral neuropathic pain (sciatica) on radicular disc conflict from disc herniation and the results obtained with oxygen-ozone treatment alone. We enrolled 318 patients with the neuroradiological diagnosis of disc herniation performed with computed tomography (CT) or magnetic resonance imaging (MRI) and symptoms characterized by low back pain complicated by sciatica, which we divided into two groups. Group A was composed of 165 patients who were treated only with oxygen-ozone therapy with CT-guided intraforaminal technique, while the remaining 153 (Group B) have undergone combined oral treatment with ALA + PEA and myrrh. Follow-up visits for the evaluation of the clinical outcome of the treatment were conducted after 60 ± 8 days using a modified version of McNab's method. At the clinical check-up, 126/165 patients included in Group A had a complete remission of pain (76.4%), while in Group B, 119/153 (77.8%) had a complete remission of pain. The results highlight how the treatment associated with ozone therapy and oral administration of alpha-lipoic acid + palmitoylethanolamide and myrrh is preferred over the simple treatment with only ozone in such patients in the phase of greatest acuity of the disease, where the pain appears to be better controlled.


Value of contrast-enhanced ultrasound in the diagnosis of breast US-BI-RADS 3 and 4 lesions with calcifications.
To evaluate the diagnostic performance of contrast-enhanced ultrasound (CEUS) for Breast Imaging-Reporting and Data System for Ultrasound (US-BI-RADS) 3 and 4 lesions with calcifications. A retrospective study of 168 breast lesions with calcifications detected on both mammography and conventional ultrasonography (US) in 152 patients were categorised as US-BI-RADS 3-4 at US between June 2009 and June 2018. CEUS scores were obtained based on a CEUS five-point scoring system. The combination of US-BI-RADS and CEUS scores created the Rerated BI-RADS (referred to as CEUS-BI-RADS). All results were compared with the histological findings. The diagnostic performances of US and CEUS-BI-RADS were compared. The diagnostic sensitivity, specificity, and accuracy of US were 81.8% (95% confidence interval [CI]: 71.6%, 92%), 85% (95% CI: 78.4%, 91.5%), and 83.9% (95% CI: 78.4%, 89.5%), respectively, while those for CEUS-BI-RADS were 98.2% (95% CI: 94.7%, 100%), 90.3% (95% CI: 84.8%, 95.7%), and 92.9% (95% CI: 89%, 96.8%), respectively. The diagnostic sensitivity and accuracy values of CEUS-BI-RADS greatly improved compared with those of US (p=0.003 and p=0.004, respectively). The areas under the receiver operating characteristic (ROC) curves for US and CEUS-BI-RADS were 0.888 (95% CI: 0.840, 0.936) and 0.963 (95% CI: 0.936, 0.989), respectively. The diagnostic efficacy of CEUS-BI-RADS was significantly higher than that of US alone (p=0.004). CEUS-BI-RADS significantly improves the diagnostic accuracy for breast US-BI-RADS 3 and 4 lesions with calcifications compared with US.


Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of murine embryonal carcinoma and embryonic stem cells.
Many pluripotent embryonal carcinoma (EC) cell lines and all embryonic stem (ES) cell lines have hitherto been maintained in the undifferentiated state only by culture on feeder layers of mitomycin C-treated embryonic fibroblasts. We now demonstrate that medium conditioned by incubation with Buffalo rat liver (BRL) cells prevents the spontaneous differentiation of such cells which occurs when they are plated in the absence of feeders. This effect is not mediated via cell selection but represents a fully reversible inhibitory action ascribed to a differentiation-inhibiting activity (DIA). BRL-conditioned medium can therefore replace feeders in the propagation of homogeneous stem cell populations. Such medium also restricts differentiation in embryoid bodies formed via aggregation of EC cells and partially inhibits retinoic acid-induced differentiation. The PSA4 EC line gives rise only to extraembryonic endoderm-like cells when aggregated or exposed to retinoic acid in BRL-conditioned medium. This suggests that DIA may be lineage-specific. DIA is a dialysable, acid-stable entity of apparent molecular weight 20,000-35,000. Its actions are reproduced neither by insulin-like growth factor-II nor by transforming growth factor-beta. DIA thus appears to be a novel factor exerting a negative control over embryonic stem cell differentiation.


Ingested manganese chloride as a contrast agent for magnetic resonance imaging.
Solutions of manganese chloride were force-fed to Sprague-Dawley rats. Magnetic resonance (MR) imaging was performed on (a) syringes containing different concentrations of manganese chloride, (b) rats after force feeding and (c) livers excised after sacrifice of the force-fed rats. Imaging was done with a 0.15-T resistive magnet. Multiple pulse sequences were used and T1 values were calculated. The signal intensity and T1 value obtained from a solution depended on the manganese concentration and the pulse sequence employed. At higher concentrations, no signal was produced due to extreme T2 shortening. Absorbed manganese affected the signal intensities and T1 values of the rats' livers. By appropriate selection of manganese concentration and pulse sequence, ingested manganese can serve as a combined gastrointestinal and hepatic MR contrast agent.


Review: Pig-based bioconversion: the use of former food products to keep nutrients in the food chain.
The primary challenge of agriculture and livestock production is to face the growing competition between food, feed, fibre, and fuel, converting them from resource-intensive to resource-efficient. A circular economy approach, using agricultural by-products/co-products, in the livestock production system would allow to reduce, reuse, and redistribute the resources. Former food products (FFPs), also named ex-foods, could represent a valid option in strengthening resilience in animal nutrition. FFPs have a promising potential to be included regularly in animal diets due to their nutritive value, although their potential in animal nutrition remains understudied. A thorough investigation of the compositional and dietary features, thus, is essential to provide new and fundamental insights to effectively reuse FFPs as upgraded products for swine nutrition. Safety aspects, such as the microbial load or the presence of packaging remnants, should be considered with caution. Here, with a holistic approach, we review several aspects of FFPs and their use as feed ingredients: the nutritional and functional evaluation, the impact of the inclusion of FFPs in pigs' diet on growth performance and welfare, and further aspects related to safety and sustainability of FFPs.


Long-term trends and spatial patterns of satellite-retrieved PM2.5 concentrations in South and Southeast Asia from 1999 to 2014.
Fine particulate matter, or PM2.5, is a serious air pollutant and has significant effects on human health, including premature death. Based on a long-term series of satellite-retrieved PM2.5 concentrations, this study analyzed the spatial and temporal characteristics of PM2.5 in South and Southeast Asia (SSEA) from 1999 to 2014 using standard deviation ellipse and trend analyses. A health risk assessment of human exposure to PM2.5 between 1999 and 2014 was then undertaken. The results show that PM2.5 concentrations increased in most areas of SSEA from 1999 to 2014 and exceeded the World Health Organization average annual limit of primary PM2.5 standards. Bangladesh, Pakistan and India experienced average PM2.5 values higher than the total average for SSEA. From 1999 to 2014, the entirety of SSEA exhibited an increased rate of 0.02μg/m3/year on average. Bangladesh and Myanmar witnessed greater incremental rates of PM2.5 than India. Correspondingly, the center of the average regional PM2.5 concentration gradually shifted to the southeast during the study period. The proportion of areas with PM2.5 concentrations exceeding 35μg/m3 increased consistently, and the areas with PM2.5 concentrations below 15μg/m3 decreased continuously. The proportion of the population exposed to high PM2.5 (above 35μg/m3) increased annually. The extent of high-health-risk areas in SSEA expanded in size and extent between 1999 and 2014, particularly in North India, Bangladesh and East Pakistan. Therefore, all of SSEA should receive special attention, and strict controls on PM2.5 concentrations in SSEA countries are urgently required.


A pretectal projection upon the accessory optic nucleus in the pigeon: an anatomical and electrophysiological study.
Neurons of the nucleus lentiformis mesencephali, the avian homologue of the optic tract nucleus, were retrogradely filled with horseradish peroxidase after injections of the enzyme into the accessory optic nucleus in pigeons. The projection appeared exclusively ipsilaterally, and was confirmed by unitary and evoked potential recordings. Together with previous reports in both birds and mammals, the present study contributes information linking the accessory optic system to the pretectal nucleus of the optic tract and thence to horizontal optomotor mechanisms.


Ethanol-induced genotoxicity.
Unrelated, nondenatured, ethanol preparations, derived synthetically or by fermentation, were found to induce qualitatively similar concentration-dependent toxic and genotoxic effects as measured by RK mutatest. In this system ethanol was found genotoxic above a threshold concentration of 18-19% (v/v) when RK+ selector cells were transiently exposed for 10 min before selection for RK- survivors at 42 degrees C.


Fetal cardiac axis in non-anomalous pregnancies: does fetal gender or maternal body mass index (BMI) matter?
To determine if cardiac axis obtained at an early ultrasound study (11-15 weeks) differs from that obtained at a late ultrasound study (18-22 weeks) in the same fetus and to evaluate the impact of fetal gender and/or maternal body mass index (BMI). Cardiac axes of 324 non-anomalous fetuses at 11-15 weeks gestation were measured, with follow-up measurements obtained at 18-22 weeks. Comparisons were performed based on gestational age period, fetal gender and obese/non-obese maternal status. (1) Mean fetal cardiac axis did not change between 11 and 15 weeks; p = 0.8, (2) mean fetal cardiac axis was more levorotated at 11-15 weeks than measurements obtained at 18-22 weeks; 48.1 ± 7.1° versus 43.7 ± 8.9°; p < 0.0001, (3) male fetuses had less levorotated cardiac axis than female fetuses in late ultrasound studies but there was no difference between them at early ultrasound studies; 18-22 weeks male fetus, 42.7 ± 9.3° versus female fetus, 45.2 ± 8.3°; p = 0.02 and 11-15 weeks male fetus, 48.1 ± 7.0° versus female fetus, 48.4 ± 7.4°, p = 0.7, respectively, and (4) similar trends with the overall study population were observed in the comparison between fetuses of obese and non-obese women. Fetal cardiac axis remains stable at 11-15 weeks, becoming less levorotated at 18-22 weeks. This may be attributed to increments in fetal lung volume. The differences in cardiac axis measurements between male and female fetuses examined at 18-22 weeks may also be attributable to differences in increment of fetal lung volume during this gestational age period.


Structural elucidation and α‑glucosidase inhibitory activity of a new xanthone glycoside from Lomatogonium rotatum (L.) Fries es Nym.
A new xanthone glycoside, 1,8-dihydroxyl-2,5-dimethoxy-xanthone-6-O-β-D-glucoside (1), along with two known xanthone glycosides and two flavonoid glycosides were isolated from the aerial parts of Lomatogonium rotatum (L.) Fries es Nym. The structure of 1 was elucidated by analysis of its spectroscopic data, including UV, IR, HR-ESI-MS and extensive 1 D and 2 D NMR techniques. In vitro test, compound 1 behaved similarity to swertianolin against α‑glucosidase and more potent inhibitory effects than the positive control, acarbose.


Establishing the European Research Area in Ageing: a network of national research programmes.
The following paper gives a summary of the European Commission's research policy with regard to the ERA-NET scheme and especially with regard to the ERA-AGE project that was launched in March 2004. All the partner institutions in the ERA-AGE project are main national funding agencies, research councils or ministries. Some selected examples of programme funding by the partner institutions may demonstrate their different ways of funding research and show the great variety of programmes. The long-term objective of the ERA-AGE project is to create a sustainable basis for the planning, implementation and exploitation of transnational research. In future, the European Research Area in Ageing could gain an added value from research investments by integrating existing funding mechanisms and research infrastructures, by commonly deciding on research priorities for funding activities in order to reduce fragmentation and duplication, and by the recruitment of young researchers.


Type A: right atrial thrombus associated with a sub-massive pulmonary embolism.
The increased use of echocardiography in patients with pulmonary thrombo-embolic disease has led to the recognition of sub-massive pulmonary embolism (SMPE) and concurrent right-heart thrombus (RHT). When these findings coincide, they are associated with a poor prognosis, identifying a high-risk group that often requires a rapid management. Since echocardiographers become the "first responders" in this clinical scenario, it is important to raise awareness and familiarize the readers regarding the importance played by the echocardiography laboratory in the prompt diagnosis and management of this entity. We describe two patients presenting with SMPE with an associated simultaneous RHT and discuss their management.


A controlled five-year follow-up study of laser trabeculoplasty as primary therapy for open-angle glaucoma.
We followed up 32 eyes of 32 patients with early glaucoma (22 with capsular glaucoma and ten with simple glaucoma) who received laser trabeculoplasty as a primary therapy. These eyes were compared with a matched control group of 32 eyes treated with medication initially. The success rate (intraocular pressure below 22 mm Hg with laser alone or medication alone) at five years was 50% (16 of 32 eyes) in the laser-treated group and 22% (seven of 32 eyes) in the control group (P less than .02). The control group required more modifications of their therapy to control intraocular pressure. The neuroretinal rim area in the control eyes decreased 2.5 times as much as in the laser group (P = .017). Changes in the Friedmann visual fields did not differ significantly between the two groups.


Pulmonary and pleural inflammation after intratracheal instillation of short single-walled and multi-walled carbon nanotubes.
Relationships between the physical properties of carbon nanotubes (CNTs) and their toxicities have been studied. However, little research has been conducted to investigate the pulmonary and pleural inflammation caused by short-fiber single-walled CNTs (SWCNTs) and multi-walled CNTs (MWCNTs). This study was performed to characterize differences in rat pulmonary and pleural inflammation caused by intratracheal instillation with doses of 0.15 or 1.5mg/kg of either short-sized SWCNTs or MWCNTs. Data from bronchoalveolar lavage fluid analysis, histopathological findings, and transcriptional profiling of rat lungs obtained over a 90-day period indicated that short SWCNTs caused persistent pulmonary inflammation. In addition, the short MWCNTs markedly impacted alveoli immediately after instillation, with the levels of pulmonary inflammation following MWCNT instillation being reduced in a time-dependent manner. MWCNT instillation induced greater levels of pleural inflammation than did short SWCNTs. SWCNTs and MWCNTs translocated in mediastinal lymph nodes were observed, suggesting that SWCNTs and MWCNTs underwent lymphatic drainage to the mediastinal lymph nodes after pleural penetration. Our results suggest that short SWCNTs and MWCNTs induced pulmonary and pleural inflammation and that they might be transported throughout the body after intratracheal instillation. The extent of changes in inflammation differed following SWCNT and MWCNT instillation in a time-dependent manner.


Considering care in context.
Sam Foster, Chief Nurse, Oxford University Hospitals, reflects on the need for regulators to take into account the care context, particularly with some trusts under more pressure than others from a second wave of COVID-19.


Bisphenol-A rapidly promotes dynamic changes in hippocampal dendritic morphology through estrogen receptor-mediated pathway by concomitant phosphorylation of NMDA receptor subunit NR2B.
Bisphenol-A (BPA) is known to be a potent endocrine disrupter. Evidence is emerging that estrogen exerts a rapid influence on hippocampal synaptic plasticity and the dendritic spine density, which requires activation of NMDA receptors. In the present study, we investigated the effects of BPA (ranging from 1 to 1000 nM), focusing on the rapid dynamic changes in dendritic filopodia and the expressions of estrogen receptor (ER) β and NMDA receptor, as well as the phosphorylation of NMDA receptor subunit NR2B in the cultured hippocampal neurons. A specific ER antagonist ICI 182,780 was used to examine the potential involvement of ERs. The results demonstrated that exposure to BPA (ranging from 10 to 1000 nM) for 30 min rapidly enhanced the motility and the density of dendritic filopodia in the cultured hippocampal neurons, as well as the phosphorylation of NR2B (pNR2B), though the expressions of NMDA receptor subunits NR1, NR2B, and ERβ were not changed. The antagonist of ERs completely inhibited the BPA-induced increases in the filopodial motility and the number of filopodia extending from dendrites. The increased pNR2B induced by BPA (100 nM) was also completely eliminated. Furthermore, BPA attenuated the effects of 17β-estradiol (17β-E₂) on the dendritic filopodia outgrowth and the expression of pNR2B when BPA was co-treated with 17β-E₂. The present results suggest that BPA, like 17β-E₂, rapidly results in the enhanced motility and density of dendritic filopodia in the cultured hippocampal neurons with the concomitant activation of NMDA receptor subunit NR2B via an ER-mediated signaling pathway. Meanwhile, BPA suppressed the enhancement effects of 17β-E₂ when it coexists with 17β-E₂. These results provided important evidence suggesting the neurotoxicity of the low levels of BPA during the early postnatal development of the brain.


Application of linear regression to videokeratoscope data for tilted surfaces.
To investigate the use of linear regression analysis performed on the tabular data display of the EyeSys videokeratoscope (VK). When a radius squared vs distance squared scatterplot is produced from aspheric surface data the equivalent conic section can be deduced from the intercept and slope of the linear regression line. Non-linear plots are often produced. Linear regression may then be applied in a number of ways. Topographical data derived from both the EyeSys VK and a computer model of the instrument were analysed by three methods of linear regression. The resultant apical radii, p-values and predicted surface tilts were compared with known values. The three methods predict different surface characteristics whose errors were found to vary depending upon the asphericity of the surface and its tilt. Apical radius is most accurately predicted by linear regression method (1). Both p-value and tilt are best predicted by averaging the radius and position data for corresponding points in each semi-meridian before squaring the resultant points and performing linear regression (method 3).


[Prolonged hospitalization versus home management for women with arrested preterm labour: a before-after study].
To compare gestational age at birth and incidence of preterm birth inferior to 37 weeks of gestation (WG) in single pregnancies with either prolonged hospitalization, or early home discharge after arrested preterm labour. Management of preterm labour was changed in our public academic hospital because no evidence of benefits for prolonged hospitalization was found and because adverse effects related to long-term hospitalization were described. In this retrospective before-after study, we compare the attitude A (December 2006 to April 2008), a prolonged hospitalization until 32 or 34WG, with new attitude B (May 2008 to February 2010), an early discharge home if the cervical exam was unchanged since admission. A total of 140 patients were included: 70 in each group. Initial hospitalization stay was significantly shorter in Group B (respectively, 5.4±5.4 days and 11.4±12.1 days; P<0.05). Preterm spontaneous delivery is 14 (20%) in group A and 21 (28.6%) in Group B (P>0.05). Gestational ages at birth were 33(5/7) and 33(0/7) WG in groups A and B, respectively (P>0.05). If we focus on patients who were discharged home in the two groups (women who did not deliver no matter the hospitalization length), the gestational age at birth (38(4/7) and 38WG; ns) and the prematurity rate inferior to 37WG (17.2% and 22.4%; ns) were statistically similar. This study suggests that shorter hospitalization does not decrease the delivery term, nor does it increase the premature delivery incidence.


Avoiding Implementation Failure in Catchment Landscapes: A Case Study in Governance of the Great Barrier Reef.
Water quality outcomes affecting Australia's Great Barrier Reef (GBR) are governed by multi-level and multi-party decision-making that influences forested and agricultural landscapes. With international concern about the GBR's declining ecological health, this paper identifies and focuses on implementation failure (primarily at catchment scale) as a systemic risk within the overall GBR governance system. There has been limited integrated analysis of the full suite of governance subdomains that often envelop defined policies, programs and delivery activities that influence water quality in the GBR. We consider how the implementation of separate purpose-specific policies and programs at catchment scale operate against well-known, robust design concepts for integrated catchment governance. We find design concerns within ten important governance subdomains that operate within GBR catchments. At a whole-of-GBR scale, we find a weak policy focus on strengthening these delivery-oriented subdomains and on effort integration across these subdomains within catchments. These governance problems when combined may contribute to failure in the implementation of major national, state and local government policies focused on improving water quality in the GBR, a lesson relevant to landscapes globally.


[Human T-lymphotropic virus (HTLV I/II) seroprevalence amongst blood-donors in a hospital Bogotá, Colombia,between 1999 and 2004].
This study was aimed at determining anti-human T-lymphotropic virus I/II (HTLV I/II) seroprevalence amongst blood-donors at the Clínica Reina Sofía in Bogotá, Colombia between 1999 and 2004. All people donating blood at the Clínica Reina Sofía were selected for anti-HTLV I/II testing; a survey was carried out which focused on risk factors. All blood donations were screened by using enzyme immunolinked assay (ELISA); repeatedly reactive serum samples were confirmed as being HTLV I or HTLV II by using Western blot (WB). 8,913 blood donors, 5,883 (66 %) males and 3,030 (34 %) females having a mean age of 37 were included in the study; 26 (0,3 %) were repeatedly reactive in ELISA tests, 6 (0,07 %) of whom were confirmed by using Western blot (WB). Our findings revealed 0,07 % HTLV seroprevalence amongst blood-donors, in line with other European and South-American countries. Although our findings suggest low HTLV I/II seroprevalence, blood-donors should be routinely screened to minimise transmission due to occult HTLV I/II infection in Colombia.


Regulation of intraflagellar transport and ciliogenesis by small G proteins.
Cilia rely on their distinctive protein compositions to function. Proteins gain access to the privileged ciliary compartment through two major routes, membrane trafficking and intraflagellar transport (IFT). Recent advances have provided two possible models for ciliary membrane transport: lateral diffusion and retention, and targeted vesicle transport. The Rab11-Rab8 cascade, which was originally discovered in the yeast's secretion pathway for bud formation, is shown to be required for cilia membrane assembly. Small GTPases, including two IFT particle subunits, and Ran, the master regulator for nuclear-cytoplasmic transport, are implicated in various aspects of IFT, a fundamental process required for the assembly of the microtubule-based backbone of cilia. This chapter reviews the key steps of ciliogenesis and possible mechanisms of IFT regulation, with emphasis on the regulatory roles of small GTPases and their regulators.