Impact of increased oxygen delivery via bovine red blood cell supplementation of culturing media on select metabolic and synthetic functions of C3A hepatocytes maintained within a hollow fiber bioreactor.
Hepatocytes are highly dependent upon appropriate oxygen provision for activity and viability. However, oxygen delivery to hepatocytes cultured within a hollow fiber bioreactor is believed to be problematic because of large diffusion distances, a high hepatocyte oxygen consumption rate and low aqueous media oxygen solubility. Supplementation of bioreactor media with bovine red blood cells (bRBCs) is one means of improving oxygen delivery to hepatocytes as hemoglobin contained within bRBCs binds oxygen. The impact of supplementing hepatocyte culturing media with bRBCs (approximately 5 x 10(8) bRBCs/ml) on hepatocyte activity (albumin and lactate production and glucose consumption) was studied. Decreased hepatocyte lactate production to glucose consumption ratios were found for the case when bRBCs were added to circulating culturing media, which indicated the presence of a more aerobic environment in comparison to the control (no bRBC supplementation). Additionally, albumin synthesis was found to be improved when the circulating media was supplemented with bRBCs. Our results thus support the use of bRBCs to improve oxygen delivery to hepatocytes maintained within a hollow fiber bioreactor.


[Urinary tract duplication].
Review the clinical and radiological characteristics of the different kinds of urinary tract duplications, assessing the outcome of the cases and the incidence of renal parenchymal injury. A retrospective study was performed including 301 children diagnosed of renal duplication by urological ultrasound, voiding cystourethrogram (VCUG) and 99-technetium renal scintigraphy (DMSA). They were classified in four groups. The first consisted of 166 (55.1%) cases that had complete ureteral duplication without an obstructive component (CD). The cases that had an upper system obstruction due to an ectopic ureter (EU), 19 (6.3%), or an ureterocele (UTC), 35 (11.6%), were divided into two different groups. The fourth group 81 (26.9%) had incomplete ureteral duplication. The nephro-urological study was performed in 181/301 after a urinary tract infection; in 100/301 after a prenatal finding of hydronephrosis. It was a casual diagnosis in 20/301. The percentage upper kidney dilation in the diagnostic ultrasound was significantly higher in those with EU 16/19 (84%) and in those with UTC 33/35 (94%), compared with the upper pole dilations found in the CD 35/166 (21%) and ID 21/81 (25%). Surgery was performed in 96/166 (41%) of the CD, 7/81 of the ID, 16/19 (84%) of the EU, and 34/35 (97%) of the UTC. Three cases with EU and 8 with UTC needed a second surgery, and 3 a third one. No scintigraphy changes were observed in 58% of the patients in the CD group, 87% in the ID group, 29% in the EU group, and 5% in the UTC group. As regards the renal duplications who did not have surgery, it was found that there were 98 refluxing units in the CD, and 74 in the ID. The spontaneous resolution of the vesicoureteral reflux (VUR) was 80% in the CD, and 90% in the ID (McNemar test P<.001). The average healing time ranged from 1 year to 5 years (Mean: 3 years and 3 months). The patients with higher risk of having renal injury are those who presented with a duplication with upper kidney obstruction due to ectopic insertion of the ureter, particularly due to an ureterocele.


Bilateral spontaneous asymptomatic urinoma: report of an unusual case.
Urinoma, defined as a collection of chronically extravasated urine, is uncommon in children. The condition is usually secondary to obstructive uropathy, commonly posterior urethral valves. Non-obstructive causes include trauma to the kidney or collecting system. Urinoma in the absence of obstruction or trauma is rare. This report describes one such boy who had bilateral spontaneous asymptomatic urinoma.


Chemical constituents from Acorus calamus with potent anti-diabetic and hepatoprotective activities.
Three previously undescribed compounds, (+)-7S,8S-syringoylglycerol-7-O-3',4'-dihydroxylphenylethanol (1), (+)-2S,3R-piscidic acid 1-methyl-5-ethyl ester (2), and 2'S-2-acetyl-3-(2,3-dihydroxypropoxyl)furan (3), together with one new natural product, 7S,8S-4,7,8-trihydroxyl-methyl phenylpropionate (4) and a known lignan (7S,8R)-methyl-4',7-epoxy-3,3'-dimethoxy-4,9-dihydroxylignan-9'-oate (5), were isolated from the ethanol extract of Acorus calamus Linn. rhizomes. Their structures were determined based on extensive spectroscopic analyses and computational methods. All the isolated compounds were evaluated for their in vitro GK activating and hepatoprotective activities, and compound 5 exhibited significant GK activating activity at 10-5 mol/L, compound 3 exhibited moderate protective effects to APAP-induced injuries of HepG2 cells at 10-5 mol/L. Furthermore, molecular docking of compound 5 bound with GK was carried out to investigate the possible structural insights into the potential binding patterns.


Erosion of X Chromosome Inactivation in Human Pluripotent Cells Initiates with XACT Coating and Depends on a Specific Heterochromatin Landscape.
Human pluripotent stem cells (hPSCs) display extensive epigenetic instability, particularly on the X chromosome. In this study, we show that, in hPSCs, the inactive X chromosome has a specific heterochromatin landscape that predisposes it to erosion of X chromosome inactivation (XCI), a process that occurs spontaneously in hPSCs. Heterochromatin remodeling and gene reactivation occur in a non-random fashion and are confined to specific H3K27me3-enriched domains, leaving H3K9me3-marked regions unaffected. Using single-cell monitoring of XCI erosion, we show that this instability only occurs in pluripotent cells. We also provide evidence that loss of XIST expression is not the primary cause of XCI instability and that gene reactivation from the inactive X (Xi) precedes loss of XIST coating. Notably, expression and coating by the long non-coding RNA XACT are early events in XCI erosion and, therefore, may play a role in mediating this process.


Center of pressure predicts Intra-limb compensatory patterns that shift demands away from knee extensors during squatting.
The persistence of knee extensor moments deficits following anterior cruciate ligament reconstruction (ACLr) may be attributed to difficulty quantifying inter- and intra-limb compensations clinically. Force plate derived center of pressure (CoPpos) and vertical force (vGRF) may provide valuable information regarding limb and joint loading impairments in this group. This study aimed to determine the: 1) relationship between measures CoPpos and intra-limb extensor moment distribution during a squat, and 2) utility of using CoPpos and vGRF to estimate knee extensor moment deficits post-ACLr. Twenty-four individuals, 142 ± 22.5 days post-ACLr, performed bilateral squats. Ankle (aEXT), knee (kEXT) and hip (hEXT) extensor moments were calculated using three-dimensional kinematics and GRF. Moments, CoPpos and vGRF were identified at peak kEXT. Intra-limb moment distribution was characterized using hEXT/kEXT and aEXT/kEXT ratios. Linear regressions analyzed relationships between CoPpos and hEXT/kEXT and aEXT/kEXT. Stepwise regressions determined if between-limb CoPpos ratio predicted between-limb ratios of hEXT/kEXT and aEXT/kEXT. Stepwise regression determined if between-limb CoPpos and vGRF ratios predicted between-limb kEXT ratio. Results found that CoPpos predicts intra-limb moment distribution (hEXT/kEXT and aEXT/kEXT); more anterior CoPpos related to higher moments at the hip and ankle relative to the knee. In addition, between-limb CoPpos ratio predicts between-limb ratio of hEXT/kEXT and aEXT/kEXT ratios. Together between-limb CoPpos (52%) and vGRF (18%) ratios explained 70% of the variance in between-limb kEXT ratios (R2 = 0.70, p < 0.001). These data suggest that force plate derived CoPpos and vGRF may be important for detection of knee extensor moment deficits in individuals post-ACLr.


DYRK3 enables secretory trafficking by maintaining the liquid-like state of ER exit sites.
The dual-specificity kinase DYRK3 controls the formation and dissolution of multiple biomolecular condensates, regulating processes including stress recovery and mitotic progression. Here, we report that DYRK3 functionally interacts with proteins associated with endoplasmic reticulum (ER) exit sites (ERESs) and that inhibition of DYRK3 perturbs the organization of the ERES-Golgi interface and secretory trafficking. DYRK3-mediated regulation of ERES depends on the N-terminal intrinsically disordered region (IDR) of the peripheral membrane protein SEC16A, which co-phase separates with ERES components to form liquid-like condensates on the surface of the ER. By modulating the liquid-like properties of ERES, we show that their physical state is essential for functional cargo trafficking through the early secretory pathway. Our findings support a mechanism whereby phosphorylation by DYRK3 and its reversal by serine-threonine phosphatases regulate the material properties of ERES to create a favorable physicochemical environment for directional membrane traffic in eukaryotic cells.


Relation between aging and circadian variation of paroxysmal atrial fibrillation.
The purpose of this study was to determine whether aging influences the circadian variation of nonvalvular paroxysmal atrial fibrillation (AF). Among 31,200 consecutive Holter monitorings recorded between January 1988 and March 1997, we detected 212 patients who had paroxysmal AF in a drug-free state. These patients were divided into 2 groups according to their age: < or = 60 years old (94 patients) and >60 years old (118 patients). In each group, the sum of the duration of each AF episode and the probability of onset, maintenance, and termination of AF were determined as hourly data and compared between the 2 groups. The time distribution of AF showed remarkable age dependence, with a well-modulated and monophasic circadian rhythm in the younger group in contrast to a toneless triphasic rhythm in the older group. Among the onset, maintenance, and termination of the arrhythmia, the most obvious age-dependence was observed in the circadian variation of onset. In the younger group, there were triple peaks with the highest one in the night, whereas the older group exhibited a single peak in the daytime. In contrast, the probabilities of maintenance and termination showed similar circadian patterns between the groups, although their amplitudes were significantly reduced in the older group. Thus, aging significantly influenced the circadian variation of paroxysmal AF, with the most prominent effect on its onset, leading to more random time-distribution of AF with increasing age. These results extend to paroxysmal AF the concept that aging disrupts rhythmicity, suggesting age-dependent differences in its pathophysiology.


Purification and physical-chemical properties of acetyl-CoA:arylamine N-acetyltransferase from pigeon liver.
Acetyl-CoA:arylamine N-acetyltransferase (EC 22.214.171.124) from pigeon liver was purified by protamine sulfate precipitation, ion exchange chromatography on DEAE-A-25 Sephadex, gel filtration on Sephadex G-75, amethopterin-AH-Sepharose 4B affinity chromatography, and finally, gel filtration on Sephadex G-100. The enzyme preparation was homogeneous as judged by ultracentrifugation studies, SDS-polyacrylamide gel electrophoresis and gel filtration. The N-terminal amino acid was detected to be histidine and the complete amino acid composition is reported. The enzyme contains one disulfide bridge and two cysteine residues/mol monomer. The isoelectric point was estimated to be 4.8. The molecular weight was determined to be 32900 by high-speed sedimentation equilibrium analysis, 33000 by Sephadex G-100 gel filtration and 31600 by SDS-disc gel electrophoresis. The sedimentation coefficient from conventional sedimentation velocity runs was 3.1 S observed by ultraviolet optics. 'Active enzyme centrifugation' showed a sedimentation constant of 5.0 and 4.8 S for the purified enzyme and crude extract from pigeon liver, respectively, indicating that the enzyme forms a dimer under conditions of catalysis. It could be demonstrated that the inhibitor amethopterin was noncompetitive with respect to the acetyl donor and the acetyl acceptor. Acetyl-CoA:arylamine N-acetyltransferase was examined in different organs of pigeon. The enzyme was not inducible by 1,3-phenylenediamine and hexobarbital in vivo.


Synthesis of Oligonucleotides containing the cis-Interstrand Crosslink Produced by Mitomycins in their Reaction with DNA.
Mitomycin C (MC) an antitumor drug and decarbamoylmitomycin C (DMC), a derivative of MC lacking the carbamoyl moiety, are DNA alkylating agents which can form DNA interstrand crosslinks (ICLs) between deoxyguanosine residues located on opposing DNA strands. MC forms primarily deoxyguanosine adducts with a 1"-R stereochemistry at the guanine-mitosene bond (1"-α, trans) whereas DMC forms mainly adducts with a 1"-S stereochemistry (1"-β, cis). The crosslinking reaction is diastereospecific: trans-crosslinks are formed exclusively at CpG sequences, while cis-crosslinks are formed only at GpC sequences. Until now, oligonucleotides containing 1"-β-deoxyguanosine adducts or ICL at a specific site could not be synthesized, thus limiting the investigation of the role played by the stereochemical configuration at C1'' in the toxicity of these compounds. Here, a novel biomimetic synthesis to access these substrates is presented. Structural proof of the adducted oligonucleotides and ICL were provided by enzymatic digestion to nucleosides, high resolution mass spectral analysis, CD spectroscopy and UV melting temperature studies. Finally, a virtual model of the 25-mer 1"-β ICL synthesized was created to explore the conformational space and structural features of the crosslinked duplex.


A Cyanide-Bridged Magnetically Switchable Cage with Encapsulated Water Molecules.
A cage complex, H2O@[Co5Fe4], was found to encapsulate two water molecules during its self-assembly. The complex exhibited remarkable multifunctionality, combining magnetic switching from the thermal electron-transfer-coupled spin transition of the cage host and dipolar reorientational motion of the confined water, as evidenced by permittivity measurements, density functional theory calculations, and solid-state 2H NMR spectra.


Continuous and Daily Oral Immunotherapy for Peanut Allergy: Results from a 2-Year Open-Label Follow-On Study.
The randomized, controlled PALISADE trial demonstrated the benefit of daily oral immunotherapy with Peanut (Arachis Hypogaea) allergen powder-dnfp (PTAH, formerly AR101) in peanut-allergic children and adolescents. ARC004, the open-label follow-on study to PALISADE, used 5 dosing cohorts to explore PTAH treatment beyond 1 year and alternative dosing regimens in peanut-allergic individuals. Active arm (PTAH-continuing) PALISADE participants who tolerated 300-mg peanut protein at the exit double-blind placebo-controlled food challenge and placebo arm (PTAH-naive) participants could enter ARC004. PTAH-continuing participants were assigned to receive daily (cohorts 1 and 3A) or non-daily (cohorts 2, 3B, and 3C) dosing regimens; PTAH-naive participants were built up to 300 mg/d PTAH, followed by maintenance dosing. At study completion, participants underwent an exit double-blind placebo-controlled food challenge with doses up to 2000 mg peanut protein. Data were assessed using descriptive statistics. Overall, 358 (87.5%) eligible participants (4-17 years) entered ARC004 (PTAH-continuing, n = 256; PTAH-naive, n = 102). Among PTAH-continuing participants, exposure-adjusted adverse event rates were 12.94 to 17.54/participant-year and 25.95 to 42.49/participant-year in daily and non-daily dosing cohorts, respectively; most participants (83%) experienced mild or moderate adverse events. Daily dosing cohorts appeared to have higher desensitization rates than non-daily dosing cohorts. Of all PTAH-continuing cohorts, cohort 3A had the longest daily dosing duration and the highest desensitization rates. Changes in immune markers with PTAH continuation demonstrated ongoing immunomodulation. Outcomes in PTAH-naive participants mirrored those of the PALISADE active arm. Continued daily PTAH treatment beyond 1 year showed sustained safety and efficacy. Ongoing immunomodulation was observed during the second year of treatment.


Early antihepatitis C virus response with second-generation C200/C22 ELISA.
Detection of early antibody to hepatitis C virus (HCV) by a new second-generation C200/C22 anti-HCV enzyme-linked immunosorbent assay (ELISA) and a four-antigen recombinant immunoblot assay (4-RIBA) was compared with the first-generation anti-HCV C100 ELISA using sequential serum samples of 9 recipients who were infected with HCV, as detected by polymerase chain reaction after transfusion of blood products. Within 26 weeks after transfusion, 9/9 (100%) recipients seroconverted with C200/22 ELISA, and 6/9 (67%) seroconverted with C100 ELISA. Compared with C100 ELISA, C200/C22 ELISA seroconversion occurred simultaneously in 3 cases, 5-6 weeks earlier in 3 other cases, and 20 weeks earlier in 1 case. Seven of 9 (78%) recipients became positive, and 2/9 (22%) became indeterminate with 4-RIBA. In 8 cases with clinical posttransfusion hepatitis non-A, non-B (PTH-NANB), anti-HCV C200/C22 ELISA seroconversion occurred 2-17 (mean 6) weeks after the onset of hepatitis. In 6 cases of PTH-NANB, anti-HCV C100 ELISA seroconversion occurred 2-26 (mean 9) weeks after the onset of hepatitis. It is concluded that the second-generation C200/C22 ELISA is more sensitive than the C100 ELISA for the detection of antibody during early HCV infection. Indeterminate 4-RIBA results are found in the early phase of HCV infection.


Capture probe conjugated to paramagnetic nanoparticles for purification of Alexandrium species (Dinophyceae) DNA from environmental samples.
To develop a rapid, cost-effective and selective Alexandrium DNA extraction procedure from environmental samples in order to provide good-quality template for the downstream PCR-based detection assay. In this study, we tested a DNA extraction method based on silica-coated, superparamagnetic nanoparticles conjugated to a DNA-capture sequence (probe) complementary to a specific region of 5.8S rDNA of the genus Alexandrium. Cultured Alexandrium catenella cells were used as the harmful algal bloom species for the DNA extraction. Then, a PCR assay was performed with primers specific for the genus Alexandrium to assess the specificity and sensitivity of the nucleic acid extraction method. This method was applied to both cultured and field samples, reaching in both cases a detection limit of one A. catenella cell. The results suggest that the use of probe-conjugated paramagnetic nanoparticles could be effective for the specific purification of microalgal DNA in cultured or environmental samples, ensuring sensitivity and specificity of the subsequent PCR assays. The DNA extraction method optimized in this study represents a progress towards the rapid and efficient direct detection of Alexandrium cells in seawater monitoring. In fact, this method requires no other equipment than a magnet and a hybridization oven and, in principle, can be adapted to different toxic microalgal species and can be automated, allowing the processing of a high number of samples.


Basal osteotomy of the first metatarsal bone in hallux valgus: experiences with the use of AO plate.
In young patients with hallux valgus, osteotomy with excision of the exostosis is frequently indicated. The basal, laterally based wedge osteotomy, is relatively seldom used. However this operation may produce a considerable reduction of the metatarsus primus varus. We modified the method by adding a small AO plate to ensure stabilization. The series consisted of 36 patients, in whom 52 feet were operated on. Pain was the most frequent indication for seeking treatment. A followup was performed an average of 1.5 years postoperatively. The subjective results were good in 41 cases, fair in nine, and poor in one. Objectively the results were good in 42 cases and fair in nine. The mean period of work was seven weeks. The metatarsus varus angle was reduced from 14.8 degrees to 9.3 degrees and the hallux valgus angle from 33.2 degrees to 17.7 degrees. The distance between the first and second metatarsal bones was reduced from 13. 7 mm to 10.2 mm. A good functional and cosmetic result was achieved in most of the cases.


Early response-based intensification of primary therapy in newly diagnosed multiple myeloma patients who are eligible for autologous stem cell transplantation: phase II study.
This phase II study prospectively evaluated the efficacy and tolerability of an early change in induction therapy before autologous stem cell transplantation (ASCT) in multiple myeloma (MM) patients who failed to achieve more than a partial response (PR) after two cycles of a cyclophosphamide, thalidomide, and dexamethasone (CTD) regimen. Patients aged 18-65 years received two cycles of CTD therapy, and then the patients who achieved more than a PR received two additional cycles of CTD therapy, while those who failed to achieve more than a PR were given intensified therapy with four cycles of a Vel-CD regimen (bortezomib, cyclophosphamide, and dexamethasone). After completing primary chemotherapy, the patients underwent ASCT. This study initially enrolled 64 patients, although four were excluded. Of the patients, 60 were treated with CTD regimen and 8 patients also had the intensified Vel-CD regimen, of whom five showing improved responses. The overall response rate before ASCT in 59 patients was 94.9 %, including 27.1 % with a stringent complete response/complete response, 23.7 % with a very good partial response (VGPR), and 44.1 % with a PR. The median time to progression (TTP) was 33.2 months (95 % CI, 26.6-34.8). Patients who attained a VGPR or better after ASCT tended to have a longer TTP than the patients who did not (not reached vs. 24.2 months, P = 0.04). In conclusion, early response-adapted intensification with a Vel-CD regimen was a well-tolerated, effective strategy for improving the response before ASCT in patients with newly diagnosed MM.


A Smart Digital Health Platform to Enable Monitoring of Quality of Life and Frailty in Older Patients with Cancer: A Mixed-Methods, Feasibility Study Protocol.
LifeChamps is an EU Horizon 2020 project that aims to create a digital platform to enable monitoring of health-related quality of life and frailty in patients with cancer over the age of 65. Our primary objective is to assess feasibility, usability, acceptability, fidelity, adherence, and safety parameters when implementing LifeChamps in routine cancer care. Secondary objectives involve evaluating preliminary signals of efficacy and cost-effectiveness indicators. This will be a mixed-methods exploratory project, involving four study sites in Greece, Spain, Sweden, and the United Kingdom. The quantitative component of LifeChamps (single-group, pre-post feasibility study) will integrate digital technologies, home-based motion sensors, self-administered questionnaires, and the electronic health record to (1) enable multimodal, real-world data collection, (2) provide patients with a coaching mobile app interface, and (3) equip healthcare professionals with an interactive, patient-monitoring dashboard. The qualitative component will determine end-user usability and acceptability via end-of-study surveys and interviews. The first patient was enrolled in the study in January 2023. Recruitment will be ongoing until the project finishes before the end of 2023. LifeChamps provides a comprehensive digital health platform to enable continuous monitoring of frailty indicators and health-related quality of life determinants in geriatric cancer care. Real-world data collection will generate "big data" sets to enable development of predictive algorithms to enable patient risk classification, identification of patients in need for a comprehensive geriatric assessment, and subsequently personalized care.


125I plaque brachytherapy for anterior uveal melanomas.
To describe the results of 125I plaque brachytherapy of uveal melanomas anterior to the equator in terms of local control and the associated complications while trying to identify their risk factors (patients' demographic data, ocular, and tumour characteristics). Retrospective analysis of a series of patients treated by 125I between 1990 and 2000 in a single institution. The main outcome measures are evaluation of local tumour control and complications associated with 125I plaque brachytherapy of these melanomas. During the study period, 136 patients were treated for an anterior tumour. The median follow-up was 62 months. The overall 5-year survival rate was 88.3%, the 5-year metastasis rate was 4% and the local recurrence rate was 1.5%. The mean final visual acuity was 20/40. The ocular complications most frequently observed at 5 years were cataract (50.3%), maculopathy (18.3%), intraocular inflammation (19.3%), and glaucoma (10.6%). Optic neuropathy, retinal detachment, keratitis, and intravitreous haemorrhage were also described. Risk factors for worse survival were age greater than 65 years and initial tumour thickness greater than 4 mm. Risk factors for the development of cataract were age more than 65 years old, male gender, and tumour diameter of more than 10 mm. Risk factors for intraocular inflammation were tumour thickness of more than 4 mm and invasion of the ciliary body. The use of 125I plaque brachytherapy to treat melanomas situated anterior to the equator allows good local and systemic control with a low rate of macular and optic disc complications. The most frequent complication was cataract formation.


Effects of prenatal betamethasone administration on leptin and adiponectin concentrations in maternal and fetal circulation.
The aim of this study was to determine the effects of in vivo administration of prenatal betamethasone on leptin and adiponectin concentration in maternal and fetal circulation. Blood samples were collected from 35 pregnant women receiving betamethasone for threatened preterm delivery before and at different time points after drug administration. Cord blood was collected at delivery in infants born from mothers treated with betamethasone and in 15 infants who delivered at the same gestational age not receiving betamethasone. Betamethasone caused an approximately 170% increase in maternal leptin at 24 hours after betamethasone, whereas it had no effects on adiponectin concentration. Betamethasone affects neonatal leptin and adiponectin levels in a time-dependent manner. The glucocorticoid-induced changes in the relationship between these adipokines in maternal and fetal circulation was long lasting. These results provide the first evidence for in vivo effects of glucocorticoids on maternal and fetal adipokines relationship in human pregnancy.


Viral Vector-Induced Ocular Hypertension in Mice.
Viral transduction of the mouse trabecular meshwork using a variety of transgenes associated with glaucoma generates an inducible and reproducible method for generating ocular hypertension due to increased aqueous humor outflow resistance of the conventional outflow pathway. Both adenovirus serotype 5 (Ad5) and lentiviruses have selective tropism for the mouse trabecular meshwork with intraocular injections. Accurate intraocular pressures are easily measured using a rebound tonometer, and aqueous humor outflow facilities can be measured in anesthetized live mice.


Photodynamic activity of thiophene-derived lysosome-specific dyes.
The photodynamic activity occurring through the lysosome photo-damage is effective in terms of triggered synergic effects which can avoid chemo-resistance pathways. The potential photodynamic activity of two fluorescent lysosome-specific probes was studied providing their interaction with human serum albumin, demonstrating their in vitro generation of singlet oxygen and investigating the resulted photo-toxic effect in human cancer cells.


In pursuit of knowledge: comparing self-explanations, concepts, and procedures as pedagogical tools.
Explaining new ideas to oneself can promote learning and transfer, but questions remain about how to maximize the pedagogical value of self-explanations. This study investigated how type of instruction affected self-explanation quality and subsequent learning outcomes for second- through fifth-grade children learning to solve mathematical equivalence problems (e.g., 7 + 3 + 9 = 7 + _ ). Experiment 1 varied whether instruction was conceptual or procedural in nature (n=40), and Experiment 2 varied whether children were prompted to self-explain after conceptual instruction (n=48). Conceptual instruction led to higher quality explanations, greater conceptual knowledge, and similar procedural knowledge compared with procedural instruction. No effect was found for self-explanation prompts. Conceptual instruction can be more efficient than procedural instruction and may make self-explanation prompts unnecessary.


GAP31 from an ancient medicinal plant exhibits anti-viral activity through targeting to Epstein-Barr virus nuclear antigen 1.
Since it was discovered as the first human tumor virus in 1964, Epstein-Barr Virus (EBV) is now implicated in several types of malignancies. Accordingly, certain aspects of EBV pathobiology have shown promise in anti-cancer research in developing virus-targeting methods for EBV-associated cancers. The unique role of EBV nuclear antigen 1 (EBNA1) in triggering episome-dependent functions has made it as the only latent gene to be expressed in most EBV+ neoplasms. Dimeric EBNA1 binds to the replication origin (oriP) to display its biological impact on EBV-driven cell transformation and maintenance. Hence, EBNA1/oriP has been made an ideal drug target site for anti-EBV protocol development. GAP31 protein was originally isolated from the seeds of an ancient medicinal plant Gelonium multiflorum. Although GAP31 has been shown to exhibit both anti-viral and anti-tumor activity, current understanding of the mechanistic picture underlying GAP31 functioning is not clear. Herein, we identify the EBNA1 DNA-binding domain as a core for GAP31 binding by performing affinity pulldown assays. Recombinant GAP31 (rGAP31) was shown to impair EBNA1-induced dimerization; consequently, it abrogated both EBNA1/oriP-mediated binding and transcription. Importantly, the therapeutic effects of GAP31 showed its capability to abrogate EBV-driven cell transformation and proliferation, and EBV-dependent tumorigenesis in xenograft animal models. Notably, the EBNA1 binding-mutant rGAP31R166A/R169A simply exhibits defective phenotypes in the above-mentioned studies. Our data suggest rGAP31 is a potential anti-viral drug which can be applied to the development of therapeutic strategies against EBV-related malignancies.


Cadmium-induced changes in avian renal morphology.
The effects of i.m. administered cadmium on growth rate and nephromorphology were studied in young pullets. The growth rate of pullets treated with 0.6 mg Cd2+/kg at 48-h intervals was severely retarded, reaching only 50% of normal growth by 21 days. Such a decrease in growth rate was prevented when cadmium was given with either ferric or magnesium EDTA chelate. Electron micrographs of kidney tissue from cadmium intoxicated birds revealed massive intracellular disorganisation of proximal tubular cells, showing increased vacuolation and dilated endoplasmic reticulum. Mitochondria were few and swollen with reduced cristae. Some disorganisation was noted in the group treated with MgEDTA in conjunction with cadmium, with normal morphology observed in the group treated with FeEDTA plus cadmium. In general, glomerular morphology of intoxicated pullets appeared normal, except that a 25% increase in thickness of the glomerular basement membrane was evident. No such membrane thickening was observed in any of the chelate treated groups. These findings indicate that both chelates can provide certain levels of protection, in terms of growth rate and morphology, from cadmium intoxication. The possible mechanisms by which chelates offer protection have been discussed, but many questions remain unanswered.


Functional connectivity alteration after real-time fMRI motor imagery training through self-regulation of activities of the right premotor cortex.
Real-time functional magnetic resonance imaging technology (real-time fMRI) is a novel method that can be used to investigate motor imagery training, it has attracted increasing attention in recent years, due to its ability to facilitate subjects in regulating the activities of specific brain regions to influence their behaviors. Lots of researchers have demonstrated that the right premotor area play critical roles during real-time fMRI motor imagery training. Thus, it has been hypothesized that modulating the activity of right premotor area may result in an alteration of the functional connectivity between the premotor area and other motor-related regions. The results indicated that the functional connectivity between the bilateral premotor area and right posterior parietal lobe significantly decreased during the imagination task. This finding is new evidence that real-time fMRI is effective and can provide a theoretical guidance for the alteration of the motor function of brain regions associated with motor imagery training.


Hydro-nium bis-(tri-fluoro-methane-sulfon-yl)amide-18-crown-6 (1/1).
The structure of the title compound, H3O+·C2F6NO4S2 -·C12H24O6 or [H3O+·C12H24O6][N(SO2CF3)2 -], which is an ionic liquid with a melting point of 341-343 K, has been determined at 113 K. The asymmetric unit consists of two crystallographically independent 18-crown-6 mol-ecules, two hydro-nium ions and two bis-(tri-fluoro-methane-sulfon-yl)amide anions; each 18-crown-6 mol-ecule complexes with a hydro-nium ion. In one 18-crown-6 mol-ecule, a part of the ring exhibits conformational disorder over two sets of sites with an occupancy ratio of 0.533 (13):0.467 (13). One hydro-nium ion is complexed with the ordered 18-crown-6 mol-ecule via O-H⋯O hydrogen bonds with H2OH⋯OC distances of 1.90 (6)-2.19 (7) Å, and the other hydro-nium ion with the disordered crown mol-ecule with distances of 1.85 (6)-2.36 (6) Å. The hydro-nium ions are also linked to the anions via O-H⋯F hydrogen bonds. The crystal studied was found to be a racemic twin with a component ratio of 0.55 (13):0.45 (13).


Correction: Smoking, alcohol use disorder and tuberculosis treatment outcomes: A dual co-morbidity burden that cannot be ignored.
[This corrects the article DOI: 10.1371/journal.pone.0220507.].


The Y chromosome: a graveyard for endogenous retroviruses.
We have isolated 20 different human endogenous retroviruses (ERV) related to ERV3, Hsrirt and Humer 4-1. Phylogeny and the presence of these ERV among different primates were determined by computer and Southern blot analyses. Preferential localization of ERV to the human, chimpanzee and orangutan Y chromosomes among the low-copy-number ERV is demonstrated. The reason for this accumulation of ERV on the strongly heterochromatic Y chromosome is probably mediated by (i) the absence of recombination of the Y chromosome that makes it more difficult for sequences to be lost, and (ii) integration of retroviruses in heterochromatic regions is less harmful to the organism. If ERV located on the Y chromosome are transcribed and translated to peptides, such peptides could be potential HY-antigens.


A solvent-template ethyl cellulose-polydimethylsiloxane crosslinking sponge for rapid and efficient oil adsorption.
Lipophilic adsorbents for oil-water separation are usually synthesized using the template method, in which hydrophobic materials are coated on a ready-made sponge. Herein, a novel solvent-template technique is used to directly synthesize a hydrophobic sponge, by crosslinking polydimethylsiloxane (PDMS) with ethyl cellulose (EC) which plays a vital role in the formation of 3D porous structure. The as-prepared sponge has advantages of strong hydrophobility, high elasticity, as well as excellent adsorption performance. In addition, the sponge can be readily decorated by nano-coatings. After the sponge was simply dipped in nanosilica, the water contact angle increases from 139.2° to 144.5°, and the maximum adsorption capacity for chiroform rises from 25.6 g/g to 35.4 g/g. The adsorption equilibrium can be reached within 3 min, and, the sponge can be regenerated by squeezing, without any change in hydrophobility or evident decline in capacity. The simulation tests of emulsion separation and oil-spill cleanup demonstrate that the sponge has great potential in oil-water separation.


Phage display of peptide epitopes from HIV-1 elicits strong cytolytic responses.
Although much effort has been expended on evaluating recombinant proteins and synthetic peptides as immunogens, they have generally proved incapable of inducing an efficient cytotoxic T-cell (CTL) response. Filamentous bacteriophage fd can display multiple copies of foreign peptides in the N-terminal region of its major coat protein pVIII, 2,700 copies of which make up the virus capsid. Here we show that fd virions displaying peptide RT2 (ILKEPVHGV), corresponding to residues 309-317 of the reverse transcriptase (RTase) of HIV-1, are able to prime a CTL response specific for this HIV-1 epitope in human cell lines. Successful priming also requires a T-helper epitope, pep23 (KDSWTVNDIQKLVGK), corresponding to residues 249-263 of HIV-1 RTase. Supplying this by displaying it on either the same or a separate bacteriophage virion led to activation of antigen-specific CD4+ T cells. Likewise, HLA-A2 transgenic mice immunized with bacteriophage virions displaying peptide RT2 were shown to mount an effective, specific anti-HIV-RT2 CTL response. This unexpected ability to elicit a designated cytolytic T-cell response, in addition to a B-cell response, has important implications for access to the class I major histocompatibility complex (MHC) loading compartment and the development of recombinant vaccines.


Observation of the inverse spin Hall effect in silicon.
The spin-orbit interaction in a solid couples the spin of an electron to its momentum. This coupling gives rise to mutual conversion between spin and charge currents: the direct and inverse spin Hall effects. The spin Hall effects have been observed in metals and semiconductors. However, the spin/charge conversion has not been realized in one of the most fundamental semiconductors, silicon, where accessing the spin Hall effects has been believed to be difficult because of its very weak spin-orbit interaction. Here we report observation of the inverse spin Hall effect in silicon at room temperature. The spin/charge current conversion efficiency, the spin Hall angle, is obtained as 0.0001 for a p-type silicon film. In spite of the small spin Hall angle, we found a clear electric voltage due to the inverse spin Hall effect in the p-Si film, demonstrating that silicon can be used as a spin-current detector.


European Central Bank׳s monetary policy decisions: A dataset of two decades of press conferences.
The dataset gathers all press conferences in text form made by the first three Presidents of the European Central Bank since its inception in 1998. Press conferences are composed of two elements: (1) introductory statements and (2) questions and answers (Q&A) from journalists. They serve as the main communication vehicle about the monetary policy decision of the ECB׳s Governing Council. From 1998 to 2016, a total of 205 press conferences have been delivered. The dataset is structured into two main sections: (1) 205 introductory statement of the Presidents of the ECB explaining the monetary policy of the Institution and (2) 205 answers provided by the Presidents to the journalists, hence a total of 410 statements (914,499 words or 3050 pages) about the monetary policy and its context, in text form.


How does our increased understanding of the role of inflammation and innate immunity in acne impact treatment approaches?
A supplement article recently published in the Journal of the European Academy of Dermatology and Venereology by Dréno et al., members of the Global Alliance to Improve Outcomes in Acne group, summarized the data for the emerging concept that inflammation in general and the innate immune system specifically play a central role in the pathogenesis of acne. This review, entitled "Understanding innate immunity and inflammation in acne: implications for management", also discusses the impact of different treatment options on the innate immune response and inflammation. The aim of the present summary is to provide a synopsis of the key points made in the paper, from the members of the Global Alliance, as relevant to the main article within this supplement: "Recent advances in the use of adapalene 0.1%/benzoyl peroxide 2.5% to treat acne patients with moderate to severe acne".


Hepatitis B virus and hepatitis C virus seroprevalence in rural areas of the southwestern region of Turkey.
Most previous studies on the frequency of hepatitis B virus (HBV) and hepatitis C virus (HCV) have analyzed data obtained from blood donors and risk groups. Few studies have been conducted in the field in rural and urban areas of Turkey. The aim of this study was to determine the seroprevalence of HBV and HCV and to investigate the association with risk factors. Between January 2006 and March 2007, 2852 people aged 18 years and over were chosen in three districts using simple random sampling, and blood samples were drawn from them. The card test technique, which is highly sensitive, was applied to blood samples for the qualitative assessment of hepatitis B surface antigen (HBsAg), anti-hepatitis B surface antigen antibodies (anti-HBs), and anti-hepatitis C virus antibodies (anti-HCV). The ELISA technique was then applied only to positive samples for confirmation. In addition, participants answered survey questions on risk factors for infection with HBV and HCV. Our results showed that 71 (2.5%) were HBsAg-positive, 462 (16.2%) were anti-HBs-positive, and 29 (1.0%) were anti-HCV-positive. Further survey results showed that seropositivity increased with some of the risk factors. Studies on seropositivity that depend on field analyses reflect the true population more accurately. We conclude that such field studies and public education activities for hepatitis B and C are essential.


Moderating role of physical activity on hippocampal iron deposition and memory outcomes in typically aging older adults.
Physical activity (PA) is linked to better cognitive and brain health, though its mechanisms are unknown. While brain iron is essential for normal function, levels increase with age and, when excessive, can cause detrimental neural effects. We examined how objectively measured PA relates to cerebral iron deposition and memory functioning in normal older adults. Sixty-eight cognitively unimpaired older adults from the UCSF Memory and Aging Center completed neuropsychological testing and brain magnetic resonance imaging, followed by 30-day Fitbit monitoring. Magnetic resonance imaging quantitative susceptibility mapping (QSM) quantified iron deposition. PA was operationalized as average daily steps. Linear regression models examined memory as a function of hippocampal QSM, PA, and their interaction. Higher bilateral hippocampal iron deposition correlated with worse memory but was not strongly related to PA. Covarying for demographics, PA moderated the relationship between bilateral hippocampal iron deposition and memory such that the negative effect of hippocampal QSM on memory performances was no longer significant above 9120 daily steps. PA may mitigate adverse iron-related pathways for memory health.


Norway rat haemoglobin phenotypes: which is the original?
On the basis of the incidence of haemoglobin alleles in different populations of rats, the hypothesis is discussed that the HbbB is phylogenetically "older".


Intestinal microsomal triglyceride transfer protein in type 2 diabetic and non-diabetic subjects: the relationship to triglyceride-rich postprandial lipoprotein composition.
Microsomal triglyceride transfer protein (MTP) is responsible for the assembly of the triglyceride-rich lipoproteins (TRLs) and is increased in diabetic animal models. Human intestinal MTP expression has not been previously reported. This study examined the relationship between intestinal MTP gene expression and postprandial TRL composition in diabetic and non-diabetic subjects. Since the MTP promoter region has a sterol response element the effect of statins on intestinal MTP mRNA was analysed. Twenty-seven diabetic and 24 non-diabetic subjects were examined. Duodenal biopsies were taken during gastroscopy and MTP mRNA was measured by RNase protection assay. Postprandial lipoprotein composition was determined. Diabetic subjects had significantly higher MTP mRNA than non-diabetic subjects. Statin therapy was associated with lower MTP mRNA in both groups. In the untreated diabetic patients compared to the untreated non-diabetic patients MTP mRNA was 25.0 +/- 25.1 amol/microg versus 13.1 +/- 5.6 amol/microg total RNA (p < 0.05). In the statin-treated diabetic group compared to statin-treated non-diabetic group MTP mRNA was 17.7 +/- 8.6 amol/microg versus 5.8 +/- 4.1 amol/microg total RNA (p < 0.05). In the whole group there was a positive correlation between the MTP mRNA and postprandial chylomicron cholesterol/B48 (r = 0.36, p < 0.01). This is the first study to demonstrate increased MTP expression in diabetic subjects. MTP mRNA expression was lower in statin-treated patients confirming the suggestion that the insulin and sterol response elements of the MTP gene are important regulators of MTP transcription in diabetes. Our results show that MTP plays a central role in regulating the cholesterol content of the chylomicron particle.


Results from Two HPV-Based Cervical Cancer Screening-Family Planning Integration Models in Malawi: A Cluster Randomized Trial.
We conducted a cluster randomized trial of two models for integrating HPV self-collection into family-planning (FP) services at 16 health facilities in Malawi between March 2020-December 2021. Model 1 involved providing only clinic-based HPV self-collection, whereas Model 2 included both clinic-based and community-based HPV self-collection. An endline household survey was performed in sampled villages and households between October-December 2021 in the catchment areas of the health facilities. We analyzed 7664 surveys from 400 villages. Participants from Model 2 areas were more likely to have ever undergone cervical cancer screening (CCS) than participants from Model 1 areas, after adjusting for district, facility location (urban versus rural), and facility size (hospital versus health center) (adjusted odds ratio = 1.73; 95% CI: 1.29, 2.33). Among participants who had ever undergone CCS, participants from Model 2 were more likely to report having undergone HPV self-collection than participants from Model 1 (50.5% versus 22.8%, p = 0.023). Participants from Model 2 were more likely to be using modern FP (adjusted odds ratio = 1.01; 95% CI: 1.41, 1.98) than Model 1 participants. The integration of FP and HPV self-collection in both the clinic and community increases CCS and modern FP uptake more than integration at the clinic-level alone.


[Expression of inducible nitric oxide synthase in human esophageal biopsies from carcinoma and precancerous lesions].
To investigate the possible role of inducible nitric oxide synthase (iNOS) in esophageal carcinogenesis. iNOS expressions in normal epithelial cells(NC), hyperplastic cells(HC), atypical hyperplastic cells(AHC) from grade I to III, carcinoma in situ(CIS), squamous cell carcinoma(SCC), and adenocarcinoma(ADC) were detected with immunohistochemistry. The positive rates of immunohistochemistry staining was 0 for NC, 2.5% for HC, 4.0%, 7.5% and 2.5% for AHC grade I to III respectively, 1.4% for CIS, 8.5% for SCC and 71.4% for ADC. There is a high expression of iNOS in human esophageal adenocarcinomas. Frequent expression of iNOS may be a potential event in ADC carcinogenesis. There is no correlation between iNOS and SCC carcinogenesis.


Detection of features associated with neovascular age-related macular degeneration in ethnically distinct data sets by an optical coherence tomography: trained deep learning algorithm.
The ability of deep learning (DL) algorithms to identify eyes with neovascular age-related macular degeneration (nAMD) from optical coherence tomography (OCT) scans has been previously established. We herewith evaluate the ability of a DL model, showing excellent performance on a Korean data set, to generalse onto an American data set despite ethnic differences. In addition, expert graders were surveyed to verify if the DL model was appropriately identifying lesions indicative of nAMD on the OCT scans. Model development data set-12 247 OCT scans from South Korea; external validation data set-91 509 OCT scans from Washington, USA. In both data sets, normal eyes or eyes with nAMD were included. After internal testing, the algorithm was sent to the University of Washington, USA, for external validation. Area under the receiver operating characteristic curve (AUC) and precision-recall curve (AUPRC) were calculated. For model explanation, saliency maps were generated using Guided GradCAM. On external validation, AUC and AUPRC remained high at 0.952 (95% CI 0.942 to 0.962) and 0.891 (95% CI 0.875 to 0.908) at the individual level. Saliency maps showed that in normal OCT scans, the fovea was the main area of interest; in nAMD OCT scans, the appropriate pathological features were areas of model interest. Survey of 10 retina specialists confirmed this. Our DL algorithm exhibited high performance for nAMD identification in a Korean population, and generalised well to an ethnically distinct, American population. The model correctly focused on the differences within the macular area to extract features associated with nAMD.


Alzheimer's Disease: Prototype of Cognitive Deterioration, Valuable Lessons to Understand Human Cognition.
It is important for neurologists to become more familiar with neuropsychological evaluation for Alzheimer disease. The growth of this method in research, as an available, inexpensive, and noninvasive diagnostic approach, which can be administered even by non-specialist-trained examiners, makes this knowledge more necessary than ever. Such knowledge has a basic role in planning national programs in primary health care systems for prevention and early detection of Alzheimer disease. This is more crucial in developing countries, which have higher rates of dementia prevalence along with cardiovascular risk factors, lack of public knowledge about dementia, and limited social support. In addition compared to the neurological hard signs which are tangible and measurable, the concept of cognition seems to be more difficult for the neurologists to evaluate and for the students to understand. Dementia in general and Alzheimer's disease as the prototype of cognitive disorders specifically, play an important role to explore all domains of human cognition through its symptomatology and neuropsychological deficits.


Accuracy of monocyte to lymphocyte ratio for tuberculosis diagnosis and its role in monitoring anti-tuberculosis treatment: Systematic review and meta-analysis.
High monocyte to lymphocyte ratio (MLR) values may be associated with the risk of active tuberculosis (TB) infection in adults, infants, and postpartum women with HIV infection. It may also serve as an indicator of the effectiveness of anti-TB treatment. Thus, the main aim of this study is to ascertain the accuracy of MLR for the diagnosis of TB and its role in monitoring the effectiveness of anti-TB therapy. This systematic review and meta-analysis followed the preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 guidelines. All statistical analyses were performed using STATA 11 and Meta-DiSc software. The Quality assessment of Diagnostic Accuracy Studies tool was used to evaluate the methodological quality of the included studies. The area under the hierarchical summary receiver-operating characteristic hierarchical summary ROC curve [(HSROC) curve (AUC)] was also calculated as an indicator of diagnostic accuracy. A total of 15 articles were included in this study. Accordingly, the result showed that elevated MLR is associated with increased risks of TB disease [odd ratio = 3.11 (95% CI: 1.40-6.93)]. The pooled sensitivity and specificity of MLR for identifying TB were 79.5% (95% CI: 68.5-87.3) and 80.2% (95% CI: 67.3-88.9), respectively. The AUC of HSROC was 0.88 (95% CI: 0.857-0.903), indicating the excellent diagnostic performance of MLR for TB. This study also showed that there is a significant reduction in the MLR value after anti-TB treatment in TB patients (standardized mean difference = 0.68; 95% CI: 0.007, 1.43). Generally, MLR can be considered as a crucial biomarker to identify TB and monitor the effectiveness of anti-TB therapy.


The pattern of neurosurgical disorders in rural northern Tanzania: a prospective hospital-based study.
The prudent allocation of neurosurgical resources and training efforts requires an understanding of prevalence and clinical pattern of neurosurgical disorders in a particular region. The aim of our study was to assess the hospital-based prevalence of neurosurgical disease in the setting of rural sub-Saharan Africa. The study was conducted at Haydom Lutheran Hospital in northern Tanzania. Over a period of 8 months, all patients suspected of having neurologic or neurosurgical disorders were seen by a neurologist in consecutive order. Patients were assessed clinically and with plain radiographs, a computed tomography scanner was not available. Of 8676 admissions, 151 patients (1.7%) were given a neurosurgical diagnosis. The most frequent diagnoses were traumatic brain injury (n=90), followed by tuberculosis of the spine (n=22), spina bifida (n=14), space-occupying cerebral lesion (n=13), and hydrocephalus (n=12). The overall mortality was 10.6%; it was especially high in patients with hydrocephalus (25%), space-occupying cerebral lesions (54%), and spina bifida (29%). Initial neurosurgical training and resource allocation in sub-Saharan Africa should focus on neurotrauma, spinal infections, congenital malformations, and hydrocephalus.


Anatomic All-Inside Anterior Cruciate Ligament Reconstruction Using Quadriceps Tendon Autograft.
All-inside anterior cruciate ligament reconstruction has recently gained popularity, in part because of its bone-sparing socket preparation and reported lower pain levels after surgery. However, because this technique uses suture loops and cortical suspension buttons for graft fixation, it has mostly been limited to looped graft constructs (e.g., hamstring autograft, peroneus longus allograft). Quadriceps tendon autograft offers several advantages in anterior cruciate ligament reconstruction but, until recently, has not been compatible with suture-loop and cortical suspensory fixation. We describe a technique that allows a relatively short (<75 mm) quadriceps tendon autograft (without bone block) to be used with established all-inside anatomic techniques.


Cytokine gene expression within the central nervous system.
1. The identification of cytokine genes expressed in the central nervous system is critical to understanding the immune network in various diseases of brain, such as infection, degeneration, and malignancy. 2. Expression of cytokine genes in human astrocytoma cell lines and in fresh brain specimens was studied by the reverse-transcribed/polymerase chain reaction method. 3. The correlation between clinical malignancy and cytokine gene expression within malignant glioma was examined, especially regarding the relevancy of inhibitory cytokines, such as transforming growth factor-beta and interleukin-10.


Subdorsal Cantilever Graft: Indications and Technique.
The subdorsal cantilever graft (SDCG) is a costal cartilage graft that is positioned below the nasal dorsum to control the position of the nasal bones and middle nasal vault. SDCG type A is used to raise the middle nasal vault and caudal nasal bones to correct the saddle nose deformity. SDCG type B can be used to raise the entire dorsum of the nose (radix, bony vault, and middle vault) in the ethnic augmentation rhinoplasty patient. This article will discuss the indications and technique of the SDCG in dorsal preservation rhinoplasty.


The post-translational trimethylation of diphthamide studied in vitro.
The amino acid diphthamide is a complex post-translational derivative of histidine that exists in eukaryotic and Archaebacterial elongation factor 2 (EF-2). Diphtheria toxin and Pseudomonas exotoxin A catalyze the transfer of an ADP-ribose residue from NAD to diphthamide, causing the inactivation of EF-2. We have used cytosolic extracts of mutant CHO-K1 cells to study the biosynthesis of diphthamide in vitro. We have identified chromatographically a precursor form of diphthamide that exists in one complementation group of mutant cells and have documented the addition of 3 methyl residues from S-adenosylmethionine to this precursor. We have identified the presence of methyltransferase capable of carrying out this reaction in vitro in cells of 15 diverse eukaryotic species.


Cross-contamination in canine and feline dietetic limited-antigen wet diets.
Adverse food reactions (AFRs) are defined as abnormal responses to an ingested food or food additive. Diagnosis and treatment of AFRs consist of the complete elimination of these ingredients in the dietary trial. Previous studies have demonstrated the presence of undeclared ingredients in commercial limited-antigen dry food diets that can compromise the results and efficacy of dietary elimination trails. The aim of this study was to assess a selection of commercial canine and feline dietetic limited-antigen wet foods for the potential cross-contamination of animal proteins from origins not mentioned on the label. Eleven canine and feline dietetic limited-antigen wet foods (9 novel animal protein foods, 1 vegetarian and 1 hydrolyzed) were analyzed by polymerase chain reaction (PCR) to detect the presence DNA of animal and vegetal origins. PCR analysis confirmed the contamination of 6 of the 11 (54.5%) limited-antigen wet diets with undeclared animal protein. One of these 6 diets was solely composed of animal protein sources completely unrelated to those declared on the label. None of the foods containing horse meat or fish were contaminated, and neither were the vegetarian or the hydrolyzed food products. Moreover, the results show that had zoological class primers only been used to check for cross-class contaminations, as are generally used in the pet food industry for in-house checks, the apparent contamination rate would have been significantly underestimated: less than 20% (3/11), instead of the actual rate of 54.7% using species-specific primers. This study reveals a high rate of cross-contamination in dietetic limited-antigen wet canine and feline foods, as previously described for dietetic dry limited-antigen foods (reported to be more than 80%). These results add new fuel to the discussion about the potential causes underlying the failure of elimination diets, since animal protein contaminants may actually be present in the commercial dietetic limited-antigen diets. AFRs may therefore occur as a result of inadequate practices in the pet food industry.


Umbilical perforation: a rare complication of entriculoperitoneal shunt.
A female baby of one-and-a-half-year old presented with extrusion of ventriculoperitoneal (VP) shunt through umbilicus. On exploration, the urachal remnant was found perforated by VP shunt and the tube exteriorized through umbilicus. Child was managed successfully with excision of urachal remnant, removal of VP shunt, antibiotic treatment and re-insertion of shunt.


[Amanita poisoning--comparison of silibinin with a combination of silibinin and penicillin].
Current treatment of amatoxin poisoning includes the administration of silibinin and penicillin in combination or silibinin alone. The aim of this study was to compare both therapeutic regimes. Of 604 patients with the suspected diagnosis of amatoxin poisoning 367 were retrospectively analysed: 118 patients had received silibinin alone and 249 patients silibinin in combination with penicillin. Logistic regression analyses were applied to investigate the efficacy of both therapeutic regimens by comparing death and liver transplantation rates. A potentially independent effect on outcome of age, sex, year of treatment, latency period of symptoms and start of silibinin therapy was taken into account. In the group who had received the combination of silibinin and penicillin 8.8% died or underwent liver transplantation compared to 5.1% in the group of those who had received silibinin alone. The risk of death or organ transplantation was thus reduced by nearly 40% in the latter group (adjusted odds ratio: 0.58; 95% CI: 0.21-1.57; p=0.28). A longer latency period (< or =12h vs. >12h) was associated with a significant reduction of this risk (adjusted OR.: 6.10; 95% CI:1.77-21.3; p=0.004). A later start of silibinin therapy (>24h vs. < or = 24h) was associated with a tendency toward an increased frequency of death or organ transplantation (adjusted OR.: 3.0; 95% CI: 0.96-9.20; p=0.059). A lower death and transplantation rate was observed in the silibinin treatment group than in group treated with silibinin combined with penicillin. However, this difference was not statistically significant. The high risk ratio relating to the time-dependent effect of silibinin suggests its efficaciousness in the treatment of amatoxin poisoning. The latency period was assessed as an independent prognostic factor.


An amino acid code for irregular and mixed protein packing.
To advance our understanding of protein tertiary structure, the development of the knob-socket model is completed in an analysis of the packing in irregular coil and turn secondary structure packing as well as between mixed secondary structure. The knob-socket model simplifies packing based on repeated patterns of two motifs: a three-residue socket for packing within secondary (2°) structure and a four-residue knob-socket for tertiary (3°) packing. For coil and turn secondary structure, knob-sockets allow identification of a correlation between amino acid composition and tertiary arrangements in space. Coil contributes almost as much as α-helices to tertiary packing. In irregular sockets, Gly, Pro, Asp, and Ser are favored, while in irregular knobs, the preference order is Arg, Asp, Pro, Asn, Thr, Leu, and Gly. Cys, His,Met, and Trp are not favored in either. In mixed packing, the knob amino acid preferences are a function of the socket that they are packing into, whereas the amino acid composition of the sockets does not depend on the secondary structure of the knob. A unique motif of a coil knob with an XYZ β-sheet socket may potentially function to inhibit β-sheet extension. In addition, analysis of the preferred crossing angles for strands within a β-sheet and mixed α-helice/β-sheet identifies canonical packing patterns useful in protein design. Lastly, the knob-socket model abstracts the complexity of protein tertiary structure into an intuitive packing surface topology map.


[Serial complement (C3 and CH 50) and immunoglobulin levels in toxaemic pregnancy (author's transl)].
The fraction of complement C3, the total complement activity and the levels of IgG, IgA and IgM immunoglobulins have been estimated in the third trimester of pregnancy in 70 control pregnant women and in 38 hypertensive pregnant women who presented either with "pure toxaemia" or with "superimposed toxaemia". The level of IgG is definitely but not significantly lowered in cases of "pure toxaemia". The other parameters that were studied showed no notable changes in comparison with control pregnant women.


Muscarinic (M1) mediation of hippocampal spontaneous theta rhythm in freely moving cats.
The effects of intrahippocampally applied different doses of muscarinic (atropine sulphate, pirenzepine, gallamine) and nicotinic (hexamethonium, mecamylamine) antagonists on the spontaneous theta rhythm in the cat hippocampal formation were investigated. The injections of atropine and pirenzepine abolished spontaneous theta but administration of gallamine did not affect the EEG pattern. The intrahippocampally administered nicotinic blockers, hexamethonium and mecamylamine, were completely ineffective in antagonizing theta waves. The data suggest that the spontaneous theta rhythm in the cat is mediated by the M1 receptor subtype.


Behavior on the water maze platform: relationship to learning and open field exploration in aged and adult rats.
In the present study we examined whether age-related deficits in the water maze could be accounted for by changes in the behavior on the platform, and whether platform behavior represents some form of exploration, akin to that seen in the open field. Twenty-seven aged and 8 adult rats (26 and 3 months old Wistar rats, respectively) were tested in an empty open field and spatial object exploration task, followed by 9 days of escape learning in a water maze. The aged as compared to the adults exhibited lower levels of open field activity and were deficient in the object displacement task. Escape deficits in the water maze and reduced activity levels on the platform were also found. In the aged, water maze performance was correlated with behavior on the platform, including quiescence, orienting-like activity and turning behavior, a form of axial rotation. In both age groups, turning behavior was also correlated with exploratory activity in the open field. In conclusion, the results support the hypothesis that age-related impairments in the water maze relate to changes in platform behavior, which, in turn, might reflect exploratory activity.


Repair of double-outlet right ventricle. An analysis of 62 cases.
Sixty-two patients with double-outlet right ventricle (DORV) underwent complete intracardiac repair between 1967 and July, 1978. Five patients (three deaths) with DORV and complete atrioventricular (AV) canal are the subject of a separate report and are not discussed further here. Twenty-eight patients had relatively uncomplicated DORV (subaortic ventricular septal defect [VSD], doubly committed VSD, or 1-malposition of the aorta with subaortic VSD) with or without pulmonary stenosis. Two (14%) of 14 died after a completely intraventricular repair, and one (12%) of eight after repair including use of a valved external conduit. Use of a transannular patch in this group was an incremental risk factor, five (83%) dying among six treated in this way. Anterior enlargement of a restrictive or unfavorably located VSD did not increase risk. A noncommitted VSD was an incremental risk factor, two (50%) of four dying after repair. No instances of complete heart block occurred in these groups. Two late deaths occurred in the uncomplicated group, from persisting servere pulmonary hypertension. The late results were good in the remainder and in the two surviving patients with noncommitted VSD. After repair of the Taussig-Bing type of DORV, eight deaths (32%) occurred among 25 patients. Complete heart block developed in two patients. In the Taussig-Bing type of DORV, the 6 year actuarial survival rate was only 38%. Most late deaths were related to improtant pulmonary vascular disease. The surgical technique that has evolved for the basic tunnel repair in the various types of DORV is described.


Retrorenal colon in pediatric patients with urolithiasis: Is the supine position for PCNL advantageous?
Percutaneous nephrolithotomy (PCNL) is one of the main approaches for the treatment of large and complex stones in pediatric patients. The patient position for PCNL has been classically divided into prone or supine and the prone position is more commonly used. One of the most feared complications of this surgical technique is colon injury and the objective of this work is to evaluate the incidence of retrorenal colon (RRC) both supine and prone position, in pediatric patients, through radiological images. A retrosepctive review was performed of all abdominal CT scans performed in one center, in patients under 18 years since 2017 to 2019. The ideal path for percutaneous puncture was traced in the upper, middle, and lower calyces of both kidneys in the prone and supine positions and contact with the kidney and adjacent organs was evaluated, with a uro-radiologist. Chi-square test and Student's t test were applied. 50 CT-scans were performed on 44 children. Patients had a mean age of 12.6 years and 71.4% were male. Twenty-five scans were performed in the prone and 25 in the supine position. The incidence of RRC for the left kidney (LK) was significantly higher in the lower calyx in prone position than in the supine position, 28% vs 4% (p = 0.049), and the right kidney (RK) presented RRC only in the prone position. The liver was the most frequent retrorenal organ (97.7%) for the RK, with a non-significant difference by position, while for the LK the spleen was the most frequent (61.1%). Colon injury is one of the most morbid complication in PCNL and one of the reasons to perform a CT scan before surgery, on which its incidence appears to be <1% in adults. The present study revealed RRC to be present in three (12%) patients in the supine position and seven (28%) patients in the prone position (p=0.15). These results were slightly higher from those reported in adults (supine: 1.7%-10%; prone: 6.8%-20%), but there are no reports in the literature on this issue to allow appropriate comparison. This is the first study to attempt to assess the incidence of RRC in children according to position. The incidence of RRC for an ideal puncture was significantly higher in the lower calyx of the left kidney in the prone position than in the supine position, and in the right kidney, the RRC only occurred in the prone position.


A Population-Based Study of Anterior Ischemic Optic Neuropathy Following Cataract Surgery.
To determine if there is an increased incidence rate of post-cataract surgery (pcs) anterior ischemic optic neuropathy (AION) compared to spontaneous AION (sAION). Retrospective, population-based cohort. Patients diagnosed with AION from January 1, 1990, through December 31, 2016, while residing in Olmsted County, Minnesota. Patients with cataract surgery preceding AION were included in the pcsAION cohort defined in 2 ways: AION within 2 months and AION within 1 year of cataract surgery. The incidence rates of pcsAION and sAION were compared using Poisson regression models. During the study period, 102 residents developed AION. The median age was 65 years (range, 40-90 years), 44 (43.1%) were female. Twenty of 102 (19.6%) patients had previous cataract surgery, of which 2 and 9 developed AION within 2 months and 1 year of surgery, respectively. The annual incidence rate of pcsAION within 2 months of surgery (8.6 per 100,000) was not significantly greater than the annual incidence rate of sAION (6.9 per 100,000; P = .78). However, the annual incidence rate of pcsAION within 1 year of surgery (38.9 per 100,000) was significantly higher than the incidence rate of sAION (6.5 per 100,000; P < .001). The incidence of AION is increased in the first year after cataract surgery, but not in the early (i.e., 2 months) postoperative period.


Which factor contributes most to the invasion resistance of native plant communities under the co-invasion of two invasive plant species?
Two invasive plant species (IPS) can co-invade the same plant community. As the number of IPS increases under the co-invasion of two IPS, plant taxonomic and functional diversity, community invasibility, community stability, invasion resistance, and invasion intensity and invasiveness of IPS and their interrelationships may be altered. This study aimed to quantify the contribution of plant taxonomic and functional diversity, community invasibility, community stability, and invasion intensity and invasiveness of IPS to the invasion resistance of native plant communities under the co-invasion of the two IPS Erigeron annuus (L.) Pers. and Solidago canadensis L. in eastern China. This study also defined a method to quantify the invasion resistance of native plant communities designated the invasion resistance index. The community-weighted mean trait values of native plants and plant diversity are the factors that are the most critical to determine the invasion resistance of native plant communities. Thus, the invasion resistance of native plant communities primarily depends on the three following factors: the relative abundance of natives, the growth performance of natives, and the diversity of natives. All levels of invasion significantly decrease the invasion resistance of native plant communities. The two IPS antagonistically affect the invasion resistance of native plant communities less under co-invasion compared with their independent invasion.


Pulsed remote Raman system for daytime measurements of mineral spectra.
A remote Raman system has been developed utilizing a 532nm pulsed laser and gated intensified charged couple device (ICCD) detector in the oblique geometry. When the system is set for 50m sample distance it is capable of measuring Raman spectra of minerals located at distances in the range of 10-65m from the telescope. Both daytime and nighttime operations are feasible and the spectra of minerals can be measured in a short period of time, of the order of a few seconds. In oblique geometry, measured sampling depth is more than 30m, during which the system maintains very high performance without any adjustments. Much longer sampling depth (0.1-120m) has been observed when the system is configured in the coaxial geometry. Clear advantages of using a gated detection mode over the continuous (CW) mode of operation in reducing the background signal and eliminating long-lived fluorescence signals from the Raman spectra are presented. The performance of the pulsed Raman system is demonstrated by measuring spectra of Raman standards including benzene (C(6)H(6)) and naphthalene (C(10)H(8)), a low Raman cross section silicate mineral muscovite (KAl(2)(Si(3)Al)O(10)(OH)(2)), and a medium Raman cross section mineral calcite (CaCO(3)).


Preparation of cellulose nanocrystals from Humulus japonicus stem and the influence of high temperature pretreatment.
As one of the most abundant wild herbs in nature, the Humulus japonicus stem (HJS) is a new low-cost source of cellulosic material. In this work, cellulose nanocrystals (CNCs) were isolated from HJS using acid hydrolysis. The influence of high temperature pretreatment (HT pretreatment) on the properties and yields of these HT-HJS (HJS fibers after HT pretreatment at different temperatures) and HT-CNCs (CNCs prepared from the HT-HJS) was studied. The results showed that there was no variation of the chemical structure among the HT-HJS and HT-CNCs. The thermal stabilities and crystallinities of the HT-HJS were higher than bleached HJS. The average diameters of the HT-CNCs were significantly smaller while the average aspect ratios of them were obviously bigger than the CNCs, and the biggest average aspect ratio of HT140-CNCs (63.40) was almost twice of CNCs (32.00). Compared with the CNCs (70.05%, 212.8°C), the crystallinity and initial degradation temperature also showed increase for the HT-CNCs and increased up to maximum of 86.93% and 227.5°C for the HT160-CNCs.


Autophagy induces mTOR-dependent glucose uptake and mTOR-independent lactate utilization in cadmium-treated A549 cells.
Cadmium (Cd) is a non-essential heavy metal with many harmful effects, especially tumorigenesis. Previously we established that autophagy-dependent increasing of glycolysis played an important role in Cd-induced cell growth and migration of A549 and HELF cells. In this study, we found Cd could induce autophagy and mTOR in A549 cells, HELF cells and in lung tissues of BALB/c mice. More interestingly, Cd-induced elevation of mTOR was autophagy-dependent and autophagy-induced cell growth and glycolysis was mTOR-dependent. However, in A549 cells, besides the above mTOR-dependent pathway, Cd-induced autophagy could directly induce PKM2 and LDHA independent of mTOR. Further study showed that only in A549 cells could autophagy other than mTOR enable Cd to increase MCT1 expression and MCT1 was involved in autophagy-induced PKM2 and LDHA. Sodium lactate added in the culture medium promoted Cd-induced cell growth of A549 cells, while had no effect on HELF cells. Finally, the effect of autophagy/MCT1/PKM2 pathway on lactate utilization to facilitate Cd-induced A549 cell growth was determined. Above all, we concluded that in HELF cells, autophagy induced mTOR-dependent glycolysis in which GLUT1 and HKII was elevated to promote glucose intake to accelerate cell growth. Whereas, in A549 cells, besides the above pathway to use glucose, autophagy could induce an mTOR-independent glycolysis pathway in which lactate could be used as fuel through autophagy-MCT1-PKM2 to escape glucose deficiency.


Low serum Α-SYNUCLEIN and oligomer Α-SYNUCLEIN levels in multiple sclerosis patients.
Multiple sclerosis (MS) is an autoimmune, inflammatory, demyelinating neurodegenerative disease progressing with attacks. Alpha-synuclein (α-Syn), a neuronal protein, has been previously associated with the inflammation and development of neurodegenerative diseases. Although the cause of neurodegeneration in multiple sclerosis is mainly associated with inflammation, α-Syn may play a role in the pathogenesis of MS, as in other classical neurodegenerative diseases such as synucleinopathies. In multiple sclerosis, α-Syn has been directly studied in central nervous system lesions and cerebrospinal fluid (CSF). However, there are few studies approaching variations in peripheral α-Syn in MS. The aim of our study was to investigate the correlation between disease progression and other clinical parameters by measuring serum α-Syn and oligomer α-Syn levels in MS patients. The study included 60 MS patients aged 18 years or older who were admitted to the Department of Neurology between 01.02.2020-01.04.2020 and diagnosed with MS according to the 2010 MC Donald criteria, and 60 age- and sex-matched healthy controls. Those who were in the MS attack period and received cortisone treatment in the past three months were excluded from the study. The serum α-Syn and oligomer α-Syn levels of the individuals in both groups were measured. The correlation between the serum α-Syn, oligomer α-Syn, oligomer α-Syn/α-Syn ratio levels of the MS patients and their age, disease duration, number of attacks, annualized relapse rate (ARR), disease type, EDSS scores and immunomodulatory drug type used was investigated. Statistical analysis was performed using the SPSS 22.0 software. In our study, 73.3% of the MS patients were female and the mean age of the patients was 36.18 ± 9.5 years. The most common MS disease type was RRMS with 83.3%. Serum α-Syn (79.52 ± 34.81) and oligomer α-Syn (18.79 ± 10.48) levels were significantly lower in the MS patients compared to the control group (p < 0.001). Serum oligomer α-Syn/α-Syn ratio was higher in the MS patients compared to the control group and in SPMS compared to RRMS, but was not statistically significant. There was no significant correlation between the serum α-Syn, oligomer α-Syn and oligomer α-Syn/α-Syn ratio ratio of the MS patients and their age, disease duration, disease type, EDDS, ARR and immunomodulatory treatments. There was a significant positive correlation between α-Syn and oligomer α-Syn in MS patients (r: 0.29, p: 0.02). In our study, serum α-Syn and oligomer α-Syn levels were lower in the MS patients compared to the control group. Low levels of α-Syn in MS may play a role in the development of neuroinflammation and may be a result of the diffuse neuronal and synaptic loss. There is a need for further studies on this subject.


"Lower abdominal pains, as if I was being squeezed…in a clamp": A Qualitative Analysis of Symptoms, Patient-Perceived Side Effects and Impacts of Ovarian Cancer.
Understanding the patient's perception of their disease is vital for guiding care decisions. The current study aimed to identify the most predominant experiences in women diagnosed with, and treated for, ovarian cancer in terms of disease-related symptoms, treatment-attributed side effects and their impacts. Semi-structured qualitative interviews about disease-related symptoms, treatment-attributed side effects and their impacts were conducted with women who were being treated for ovarian cancer in Europe (n = 55) or in the USA (n = 9). The women were also asked to rate the bothersomeness of the symptoms, side effects and impacts that they mentioned during the interview. Symptoms, side effects and impacts were identified from coded interview transcripts using an iterative coding framework. Bloating, abdominal pain, tiredness and frequent urination were the most frequently expressed symptoms, and were reported by 72%, 67%, 64% and 55% of women, respectively, which together constituted approximately 30% of all symptom expressions. The most bothersome symptoms were reported as bloating, abdominal pain, pain in the side, tiredness and fatigue. The most frequently expressed side effects were hair loss, neuropathy, tiredness and nausea, which were reported by 84%, 63%, 61% and 61% of women, respectively. The most bothersome reported side effects were constipation, nausea, diarrhoea, pain in general, fatigue, weakness, reduced sleep quality and hair loss. Feelings of anxiety, concerns about the future, physical functioning, work limitations and the adoption of coping strategies were the most frequently expressed impacts and were reported by 72-80% of women. Impacts reported as the most difficult to deal with were concerns about the future, emotional difficulties in general, physical functioning, sexual functioning, negative self-image, fatigue, sleep difficulties, financial burden and work limitations. In our qualitative study, the most common and most bothersome experiences reported by women treated for ovarian cancer were symptoms of bloating, abdominal pain and tiredness; side effects of hair loss, nausea and tiredness/fatigue; and impacts relating to concerns about the future, physical functioning and work limitations. We suggest that clinicians measure these experiences consistently and take them into consideration when making treatment decisions.


Effect of fiber from fruits and vegetables on metabolic responses of human subjects I. Bowel transit time, number of defecations, fecal weight, urinary excretions of energy and nitrogen and apparent digestibilities of energy, nitrogen, and fat.
Twelve men 37 to 58 years of age consumed two diets for a period of 26 days each in a cross-over design. The high fiber diet contained fruits and vegetables and the lower fiber diet contained fruit and vegetable juices. Neither diet contained whole grain cereals or nuts. Some nutrients were added to the low fiber diet in order to make the diets equivalent insofar as possible in all respects except fiber. The inclusion of fruits and vegetables in the diet decreased bowel transit time, increased fecal weight, increased number of defecations, increased fecal excretions and decreased apparent digestibilities of energy, nitrogen, and fat. Six of the 12 subjects had diastolic blood pressure of 80 or more when consuming the low fiber diet. The diastolic blood pressures of these six men were significantly lower when the high fiber diet was consumed.


Preparation, characterization and in vitro release of gentamicin from PHBV/wollastonite composite microspheres.
Composite microspheres have been prepared from bioactive wollastonite (W) and biodegradable poly (hydroxybutyrate-polyhydroxyvalerate) (PHBV) in the present study. Gentamicin was encapsulated into the microspheres by the absorption method and the in vitro release of the gentamicin from the microspheres was performed in distilled water, modified simulated body fluid (SBF) and phosphate buffered saline (PBS) at 37 degrees C for 22 days, respectively. The results showed that the release behavior of gentamicin from PHBV/W composite microspheres was similar to that from the pure PHBV microspheres when the experiment was performed in distilled water. However, in the PBS and SBF solutions, gentamicin released from the PHBV/W composite microspheres at a relatively lower rate as compared to that of the pure PHBV microspheres and 90% of the total amount of gentamicin released from the composite microspheres after soaking for 22 days, which was much longer than that for the release of the same amount gentamicin from the pure PHBV microspheres (8 days). Scanning electron microscopy (SEM) and energy-dispersive spectrometer (EDS) analysis on the microspheres after release in SBF and PBS revealed that a microporous apatite layer was formed on the composite microspheres surface, which resulted in a controlled release behavior of the gentamicin from the PHBV/W composite microspheres. All of these results provided the possibility that the PHBV/W composite microspheres could be applied as alternative drug controlled release systems, especially as bone fillings for bone repair due to their advantages of controlled releasing antibiotics and apatite-formation ability, through which the implanted microspheres could chemically bond to the surrounding tissue in vivo.


[Usefulness of video-assisted cervicotomy on the extraction of foreign bodies].
Foreign bodies located in the upper aerodigestive tract, if not diagnosed and taken out speedily, may be the cause of great morbimortality. We present a female patient who arrived at the emergency department reporting she had swallowed a fish bone; it was finally located in the pre-vertebral muscle at the level of C3-C4, where it had entered from the oesophagus at 2 cm from the superior oesophageal sphincter. We used a cervicotomy assisted by endoscopy, showing a new application for videoendoscopy.


Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1.
Human cytochrome P450 46A1 (CYP46A1) is one of the key enzymes in cholesterol homeostasis in the brain. The crystallization and heavy-atom structure solution of an active truncated CYP46A1 in complex with the high-affinity substrate analogue cholesterol-3-sulfate (CH-3S) is reported. The 2.6 angstroms structure of CYP46A1-CH-3S was solved using both anion and cation heavy-atom salts. In addition to the native anomalous signal from the haem iron, an NaI anion halide salt derivative and a complementary CsCl alkali-metal cation salt derivative were used. The general implications of the use of halide and alkali-metal quick soaks are discussed. The importance of using isoionic strength buffers, the titration of heavy-atom salts into different ionic species and the role of concentration are considered. It was observed that cation/anion-binding sites will occasionally overlap, which could negatively impact upon mixed RbBr soaks used for multiple anomalous scatterer MAD (MMAD). The use of complementary cation and anion heavy-atom salt derivatives is a convenient and powerful tool for MIR(AS) structure solution.


Proconvulsant and anticonvulsant effects in mice of acute and chronic treatment with cocaine.
The proconvulsant and anticonvulsant effects of acute and chronic exposure to cocaine were investigated in adult, male, CF-1 mice. The proconvulsant effects of cocaine in mice only manifested themselves after daily exposure to motor-stimulant doses. Although daily treatment decreased electroshock convulsion threshold, no motor convulsions were observed. Animals in the proconvulsant state, however, kindled to electrically-induced convulsions more rapidly than did controls. Furthermore, daily treatment with cocaine and electroshock also enhanced the development of electrical kindling. These results illustrate that the excitatory properties of cocaine in the CNS can enhance phenomena which cause a persistent increase in excitability of the CNS. In contrast to the proconvulsant activity after chronic exposure, cocaine, administered acutely, in motor-stimulant doses, was anticonvulsant in a variety of tests using electroshock and chemically-induced convulsions. The drug elevated electroshock thresholds for both minimal and maximal convulsions and these responses were not blocked by haloperidol. In tests for minimal chemically-induced convulsions, cocaine elevated the threshold to N-methyl-DL-aspartate, but not to bicuculline; against maximal convulsions, the drug was anticonvulsant against both N-methyl-DL-aspartate and bicuculline. Cocaine did not affect convulsion thresholds for strychnine, arecoline or aminophylline; these data suggest that the anticonvulsant action of cocaine is relatively selective for the gamma-aminobutyric acid (GABA) and glutamate systems.


Speed-controlled venovenous modified ultrafiltration for pediatric open heart operations.
Although modified ultrafiltration after pediatric open heart operations is used by several clinical centers, the risk of complications is a matter of concern. This report describes a simple, reliable, and reproducible technique of speed-controlled venovenous modified ultrafiltration.


Ubiquitin-Proteasome Axis, Especially Ubiquitin-Specific Protease-17 (USP17) Gene Family, is a Potential Target for Epithelial-Mesenchymal Transition in High-Grade Serous Ovarian Cancer.
To investigate gene expression differences and related functions between primary tumor, malignant cells in ascites, and metastatic peritoneal implant in high-grade serous ovarian cancer. Biopsies from primary tumor, peritoneal implant, and ascites were collected from 10 patients operated primarily for high-grade, advanced-staged serous ovarian cancer. Total RNA isolation was performed from collected tissue biopsy and fluid samples, and RNA expression profile was measured. Messenger RNA expression profiles of 3 different groups were compared. Functional analyses of candidate genes were carried out by gene ontology and pathway analysis. There were significant differences in the expression of 5 genes between primary tumor and peritoneal implant, 979 genes between primary tumor and malignant cells in ascites, and 649 genes between peritoneal implant and malignant cells in ascites. Three commonly enriched gene ontology functions between "primary tumor and malignant cells in the ascites" and "peritoneal implant and malignant cells in the ascites" were protein deubiquitination, ubiquitin-dependent protein catabolism, and apoptotic processes. All genes related to these functions belonged to USP17 gene family. Gene expression difference between primary tumor and the peritoneal implant is not as much as the difference between primary tumor and free cells in the ascites. These results show that malignant cells in the ascites return into its genetic origin after they invade on the peritoneum. Significantly increased expression of DUB-enzyme genes, SNAR gene family, and ribosomal pathway genes in epithelial-mesenchymal transition suggests that this regulation is ubiquitin-proteasome dependent. Especially, this is the first study that offers USP17 as a potential target for epithelial-mesenchymal transition.


The Yugoslavia Prospective Study of environmental lead exposure.
The Yugoslavia Prospective Study of environmental lead exposure has studied the associations between exposure to lead and pregnancy outcomes; childhood neuropsychological, behavioral, and physical development; and hematologic, renal, and cardiovascular function. The cohort comprises 577 children born to women recruited at midpregnancy in two towns in Kosovo, Yugoslavia; one town is the site of a lead smelter, refinery, and battery plant and the other is 25 miles away and relatively unexposed. A sample of these children has been followed at 6-month intervals through 7.5 years of age. Blood lead concentrations ranged from 1 to 70 microg/dl. Exposure to lead was not associated with adverse pregnancy outcomes. Exposure was associated with modest decrements in intelligence, small increases in blood pressure, higher risks of proteinuria, small increases in behavior problems, and perturbed hematopoiesis. Only at low level exposures (i.e., <16 microg/dl) were small associations with decreased height found. We discuss methodological problems that may hinder causal interpretation of these data, namely, use of blood lead concentration as an exposure measure, confounding, and town-specific associations. We conclude that while reported associations are small, collectively they lend support to the notion that lead is a toxicant with numerous adverse health effects.


Addition of polyglycol to old culture of Phanerochaete chrysosporium.
Polyglycols increased lignin peroxidase activity in shaken cultures of Phanerochaete chrysosporium even when they were added to an old fungus (5 or 10 d after inoculation). The effects depended on the polyglycol mol wts (10(2)-10(6) Daltons) as well as on the backbone structure (i.e., poly[ethylene glycol], poly[butylene glycol], poly[propylene glycol]) and terminal groups (i.e., poly[ethylene glycol], poly[ethylene glycol] methyl ether, poly[ethylene glycol] dimethyl ether). The residual quantity of polyglycol in the biomass and in the culture filtrate also varied among different polyglycols. The polyglycols act after being adsorbed to the cell membrane exaggerating the asymmetry of the membrane environment. In old fungus, the incompatibility of polyglycols and glycans decreases the adsorption and the effect of high mol wt polyglycols.


A meta-analysis on the toxicity of microcystin-LR to fish and mammals.
Microcystin-leucine arginine (MC-LR), the most prevalent and dangerous microcystin, poses high risks to living organisms, especially fish and mammals. Although many studies have focused on the toxic effect on fish and mammals exposed to MC-LR, works that incorporate published data into a comprehensive comparison and analysis are still limited. Here, the adverse effects of oxidative stress markers, health, functional traits, and performance traits in fish and mammals were systematically verified by collecting data from 67 studies for the first time. Notably, we first found that the activities of malondialdehyde (MDA) (p < 0.05) and lactoperoxidase (LPO) always showed increases, whereas the growth (performance traits) always had a significant decrease (p < 0.001) under all variables of MC-LR exposure, i.e., exposure time, exposure concentration, exposure route, and even life stage. Additionally, our study first verified that the activities of MDA and LPO can be employed as oxidative stress indicators of MC-LR effects in fish and mammals instead of other biomarkers of oxidative stress, such as superoxide dismutase (SOD) and catalase (CAT), considered by previous studies. Growth may be regarded as a highly sensitive indicator of MC-LR toxicity in mammals and fish. At the same time, we first found that the impact of MC-LR exposure concentration on LPO, MDA, and growth is higher than that of exposure time, exposure route, and different life stages using the random forest (RF) model. In short, this work sheds light on the potential biochemical and individual toxicity of MC-LR exposure in fish and mammals.


Complete genome sequence of an extensively drug-resistant Pseudomonas aeruginosa ST773 clinical isolate from North India.
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen causing a wide range of community- and hospital-acquired infections. Here we report the complete genome sequence of an extensively drug-resistant (XDR) P. aeruginosa strain (PA790) in order to understand the antibiotic resistance genes (ARGs) harboured by such a strain. Whole-genome sequencing (WGS) was performed using Illumina HiSeq and Nanopore MinION platforms. Genome assembly was performed using Unicycler v.0.4.8. The genome was annotated using the NCBI Prokaryotic Genome Annotation Pipeline (PGAP). In silico predictions were fulfilled using curated bioinformatics tools. Pseudomonas aeruginosa PA790 was classified as XDR and belongs to sequence type 773 (ST773). The complete genome size is 6 932 250 bp with a G+C content of 66.02% and a BUSCO (Benchmarking Universal Single-Copy Orthologs) score of 100. Strain PA790 harboured 12 different ARGs conferring resistance to eight different classes of antibiotics. It was identified as the nineteenth ST773 strain among 5785 whole-genome sequences of P. aeruginosa available in the NCBI database. Pseudomonas aeruginosa PA790 belongs to ST773 and was identified as the nineteenth such isolate to be submitted to NCBI and the first complete ST773 genome from India. The WGS data with multiple ARGs of P. aeruginosa PA790 (ST773) will aid in understanding the evolution and phylogeny of such high-risk clones and provide a solid basis for further research on XDR strains.


Icariin-induced inhibition of SIRT6/NF-κB triggers redox mediated apoptosis and enhances anti-tumor immunity in triple-negative breast cancer.
Abnormal activation of the nuclear factor-kappa B (NF-κB) signaling pathway is closely implicated in triple-negative breast cancer growth, metastasis, and tumor immune escape. In this study, the anti-cancer effects of icariin, a natural flavonol glycoside, toward breast cancer cells and the underlying mechanisms were investigated. This investigation showed that icariin selectively inhibited proliferation and triggered apoptosis in breast cancer cells in a concentration- and time-dependent manner, but exhibited little cytotoxicity in normal breast cells. Moreover, icariin induced cell apoptosis via a mitochondria-mediated pathway, as indicated by the upregulated ratio of Bax/Bcl-2 and reactive oxygen species induction. Importantly, icariin impaired the activation of the NF-κB/EMT pathway, as evidenced by upregulation of SIRT6, resulting in inhibition of migration and invasion of breast cancer cells. Additionally, oss-128167, an inhibitor of SIRT6, dramatically attenuated anti-migration and anti-invasion effects of icariin. Transcriptomic analysis verified that impairment of NF-κB led to the selective function of icariin in breast cancer cells. Notably, icariin exhibited a significant tumor growth inhibition and anti-pulmonary metastasis effect in a tumor mouse model of MDA-MB-231 and 4T1 cells by regulating the tumor immunosuppressive microenvironment. Together, these results showed that icariin could effectively trigger apoptosis and inhibit the migration of breast cancer cells via the SIRT6/NF-κB signaling pathway, suggesting that icariin might serve as a potential candidate drug for the treatment of breast cancer.