Changes in serum thyroid hormones levels and their mechanisms during long-term growth hormone (GH) replacement therapy in GH deficient children.
The effects of GH therapy on thyroid function among previous reports have shown remarkable discrepancies, probably due to differences in hormone assay methods, degree of purification of former pituitary-derived GH preparations, dosage schedules, diagnostic criteria, patient selection, duration of treatment and study design. These considerations motivated us to investigate whether and how GH replacement therapy changes serum thyroid hormone levels, including the much less studied rT3 levels, in a group of unequivocally GH-deficient children receiving long-term recombinant human GH therapy. Twenty clinically and biochemically euthyroid children were studied in two therapeutic conditions: on GH replacement therapy for at least 6 months and without GH replacement, either before GH was started or after GH was withdrawn for 30-60 days. Eight patients were on thyroxine replacement treatment and thyroxine doses were kept constant during the study. Blood was collected before and after 15, 20 and 60 minutes of TRH administration in both therapeutic conditions (with GH and without GH). Concentrations of thyroid hormone levels were determined only in sera obtained before TRH administration. FT4, T3 and TSH were measured by immunoflourimetric assays and rT2 was measured by immunoradioassay. Patients were classified into two groups, according to basal TSH levels: group I (TSH > 0.4 mU/l, n = 12) and group II (on thyroxine and TSH < 0.05 mU/l, n = 8). In both groups, serum FT4 levels decreased (17. 0 +/- 1.1 vs. 14.3 +/- 0.9 mU/l, P < 0.001, and 18.0 +/- 1.7 vs. 14. 2 +/- 1.7 mU/l, P < 0.01, respectively), serum T3 levels increased (1.8 +/- 0.1 vs. 2.4 +/- 0.2 nmol/l, P < 0.001, and 1.9 +/- 0.3 vs. 2.4 +/- 0.2 nmol/l, P < 0.05, respectively), and serum rT3 levels decreased (0.35 +/- 0.03 vs. 0.25 +/- 0.03 nmol/l, P < 0.01, and 0. 48 +/- 0.06 vs. 0.34 +/- 0.06 nmol/l, P < 0.01, respectively). Basal (3.2 +/- 0.50 vs. 2.6 +/- 0.72 mU/l, P = 0.28, paired t-test), TRH-stimulated peak TSH levels (13.9 +/- 5.3 vs. 15.9 +/- 8.0 mU/l, P = 0.35, paired t-test) and TRH-stimulated TSH secretion, expressed as area under the curve (609 +/- 97 vs. 499 +/- 53 mU/l.minutes-1, P = 0.15, paired t-test), remained unchanged during GH replacement in group I patients. Low serum FT4 and high serum T3 levels were observed in only one patient each, but low serum rT3 levels were found in six patients (four in group I and two in group II) during GH replacement. These results show that long-term GH replacement therapy in children with unequivocal GHD significantly decreases serum FT4 and rT3 levels and increases serum T3 levels; that these changes are independent of TSH and result from increased peripheral conversion of T4 to T3 and that GH replacement therapy in GH deficient children does not induce hypothyroidism, but simply reveals previously unrecognized cases whose serum FT4 values fall in the low range during GH replacement.


Plasticity and regulatory mechanisms of human ILC2 functions.
Human group 2 innate lymphoid cells (ILC2) represent the innate counterpart of Th2 cells and cooperate with them in helminths protection and in the pathogenesis of allergic diseases. Some reports described ILC2 plasticity and few studies investigated the cellular and molecular mechanisms regulating human ILC2 functions. The aim of this study is to define how immune deviation and immune regulation control human ILC2-mediated immune response. Human circulating ILC2 were expanded in vitro and then cultured in presence of IL-12 or IL-1β plus IL-23 or co-coltured in presence of circulating CD4+CD25highFoxp3+Treg. IL-12 induces IFN-γ production and upregulation of T-bet mRNA level on human circulating ILC2 whereas IL-1β and IL-23 mediate IL-22 production and upregulation of RORC mRNA level. In all these conditions, GATA-3 mRNA level is not reduced and the typical type 2 cytokines are only partially reduced. Moreover, "modulated" ILC2 have reduced ability to induce IgE producing by B cells. ILC2 proliferation, cytokines production and CD154 expression were inhibited by CD4+CD25highFoxp3+ Treg cells. TGF-β reduced CD154 expression on ILC2 stimulated with IL-25/IL-33. This study defines possible cellular and molecular mechanisms responsible for modulation and inhibition of human ILC2 activity. These results may be useful in the development of strategies aimed to dampen ILC2 function in type-2 mediated diseases.


Caesarean section. A clinical study with special reference to the increasing section rate.
At Huddinge University Hospital 539 Caesarean Sections (C.S.) were made among 8415 deliveries from October 1972 to June 1976, corresponding to an over all C.S. rate of 6.4%. Over these years the rate has increased from 3.5% in 1972 to 9.7% in 1976. The main increase was due to a higher incidence of abdominal deliveries in cases of imminent fetal asphyxia. The maternal complication rate and the neonatal morbidity rate were both 6.5 times higher and the neonatal mortality rate was 4.1 times higher in emergency than in elective surgery. There was neither any mortality nor any morbidity in infants delivered by elective C.S. from healthy mothers at term.


O-Acetylhomoserine Sulfhydrylase from Clostridioides difficile: Role of Tyrosine Residues in the Active Site.
O-acetylhomoserine sulfhydrylase is one of the key enzymes in biosynthesis of methionine in Clostridioides difficile. The mechanism of γ-substitution reaction of O-acetyl-L-homoserine catalyzed by this enzyme is the least studied among the pyridoxal-5'-phosphate-dependent enzymes involved in metabolism of cysteine and methionine. To clarify the role of active site residues Tyr52 and Tyr107, four mutant forms of the enzyme with replacements of these residues with phenylalanine and alanine were generated. Catalytic and spectral properties of the mutant forms were investigated. The rate of γ-substitution reaction catalyzed by the mutant forms with replaced Tyr52 residue decreased by more than three orders of magnitude compared to the wild-type enzyme. The Tyr107Phe and Tyr107Ala mutant forms practically did not catalyze this reaction. Replacements of the Tyr52 and Tyr107 residues led to the decrease in affinity of apoenzyme to coenzyme by three orders of magnitude and changes in the ionic state of the internal aldimine of the enzyme. The obtained results allowed us to assume that Tyr52 is involved in ensuring optimal position of the catalytic coenzyme-binding lysine residue at the stages of C-α-proton elimination and elimination of the side group of the substrate. Tyr107 could act as a general acid catalyst at the stage of acetate elimination.


[Orthoses for rheumatic hand deformities].
Treatment of rheumatic hand deformities is a very sensitive task, requiring profound knowledge of functional anatomy as well as extensive rheumatologic experience. Assistive devices such as orthoses in the therapy of typical deformities aim to prevent continuing loss of function. Furthermore reduction of pain, economization of movements and maintaining postoperative results are important. Contraindications for surgical interventions are also an indication. Static orthoses are to be distinguished from individually produced dynamic devices, which may protect anatomical structures while providing relatively free motion. The indication for this treatment must be agreed by the physician in charge, the patient and the occupational therapist together, assisting functional exercises are strongly recommended. The condition of the skin must be considered and regular follow-up has to be arranged. Each region of the hand and characteristic deformities require individual orthotic devices, according to the capabilities of the patient. The patient's possible improvement of daily activities in relation to the disadvantages of having to use orthoses determines the therapy. Given indications for surgical procedures are taken into consideration from the onset of the treatment plan.


Performance evaluation of machine learning-based infectious screening flags on the HORIBA Medical Yumizen H550 Haematology Analyzer for vivax malaria and dengue fever.
Automated detection of malaria and dengue infection has been actively researched for more than two decades. Although many improvements have been achieved, these solutions remain too expensive for most laboratories and clinics in developing countries. The low range HORIBA Medical Haematology Analyzer, Yumizen H550, now provides dedicated flags 'vivax malaria' and 'dengue fever' in routine blood testing, developed through machine learning methods, to be used as a screening tool for malaria and dengue fever in endemic areas. This study sought to evaluate the effectiveness of these flags under real clinical conditions. A total of 1420 samples were tested using the Yumizen H550 Haematology Analyzer, including 1339 samples from febrile patients among whom 202 were infected with malaria parasites (Plasmodium vivax only: 182, Plasmodium falciparum only: 18, both: 2), 210 were from febrile dengue infected patients, 3 were from afebrile dengue infected patients and 78 were samples from healthy controls, in an outpatient laboratory clinic in Mumbai, India. Microscopic examination was carried out as the confirmatory reference method for detection of malarial parasite, species identification and assessing parasitaemia based on different stages of parasite life cycle. Rapid diagnostic malarial antigen tests were used for additional confirmation. For dengue infection, NS1 antigen detection by ELISA was used as a diagnostic marker. For the automated vivax malaria flag, the original manufacturer's cut off yielded a sensitivity and specificity of 65.2% and 98.9% respectively with the ROC AUC of 0.9. After optimization of cut-off value, flag performance improved to 72% for sensitivity and 97.9% specificity. Additionally it demonstrated a positive correlation with increasing levels of parasitaemia. For the automated dengue fever flag it yielded a ROC AUC of 0.82 with 79.3% sensitivity and 71.5% specificity. The results demonstrate a possibility of the effective use of automated infectious flags for screening vivax malaria and dengue infection in a clinical setting.


Isoform and tissue dependent impact of apolipoprotein E on adipose tissue metabolic activation: The role of apolipoprotein A1.
Adipose organ is made of white (WAT) and brown (BAT) adipose tissue which are primarily responsible for lipid storage and energy production (heat and ATP) respectively. Metabolic activation of WAT may ascribe to this tissue characteristics of BAT, namely non-shivering thermogenesis and ATP production. Recent data indicate that apolipoproteins E (APOE) and A1 (APOA1) regulate WAT mitochondrial metabolic activation. Here, we investigated the functional cross-talk between natural human APOE2 and APOE4 isoforms with APOA1 in this process, using Apoe2knock-in and Apoe4knock-in mice. At baseline when Apoe2knock-in and Apoe4knock-in mice express both APOE and Apoa1, the Apoe2knock-in strain appears to have higher mitochondrial oxidative phosphorylation levels and non-shivering thermogenesis in WAT compared to Apoe4knock-in mice. When mice were switched to a high-fat diet for 18 weeks, circulating levels of endogenous Apoa1 in Apoe2knock-in mice became barely detectable though significant levels of APOE2 were still present. This change was accompanied by a significant reduction in WAT mitochondrial Ucp1 expression while BAT Ucp1 was unaffected. Ectopic APOA1 expression in Apoe2knock-in animals potently stimulated WAT but not BAT mitochondrial Ucp1 expression providing further evidence that APOA1 potently stimulates WAT non-shivering thermogenesis in the presence of APOE2. Ectopic expression of APOA1 in Apoe4knock-in mice stimulated BAT but no WAT mitochondrial Ucp1 levels, suggesting that in the presence of APOE4, APOA1 is a trigger of BAT non-shivering thermogenesis. Overall, our data identified a tissue-specific role of the natural human APOE2 and APOE4 isoforms in WAT- and BAT-metabolic activation respectively, that appears dependent on circulating APOA1 levels.


Natural and artificial radioactive pollution in sediment and soil samples of the Bosphorus, Istanbul.
This study focused on the determination of natural (238U, 232Th, and 40K) and artificial (137Cs) radionuclide concentrations both in 55 sediment samples collected from various depths in the Bosphorus and 5 soil samples from the coastline of the Bosphorus, Istanbul, using gamma-ray spectrometry with an HPGe detector. The mean activity concentrations of natural 238U, 232Th, and 40K and anthropogenic 137Cs were determined to be 11.41 ± 0.21 Bq kg-1, 6.87 ± 0.16 Bq kg-1, 369.61 ± 3.41 Bq kg-1, and 6.54 ± 0.11 Bq kg-1, respectively, in the sediment samples. The average activity concentrations of 238U, 232Th, 40K, and 137Cs radionuclides in the soil samples were also measured to be 11.65 ± 0.18 Bq kg-1, 9.55 ± 0.15 Bq kg-1, 369.43 ± 3.09 Bq kg-1, and 4.57 ± 0.09 Bq kg-1, respectively. Radiological contour maps based on the activity concentrations of natural and artificial radionuclides in the sediment samples for the Bosphorus, Istanbul, were created. The total annual effective doses due to soil samples were calculated to be 34.58 μSv y-1.


Helical self-assembly of a mucin segment suggests an evolutionary origin for von Willebrand factor tubules.
The glycoprotein von Willebrand factor (VWF) contributes to hemostasis by stanching injuries in blood vessel walls. A distinctive feature of VWF is its assembly into long, helical tubules in endothelial cells prior to secretion. When VWF is released into the bloodstream, these tubules unfurl to release linear polymers that bind subendothelial collagen at wound sites, recruit platelets, and initiate the clotting cascade. VWF evolved from gel-forming mucins, the polymeric glycoproteins that coat and protect exposed epithelia. Despite the divergent function of VWF in blood vessel repair, sequence conservation and shared domain organization imply that VWF retained key aspects of the mucin bioassembly mechanism. Here, we show using cryo-electron microscopy that the ability to form tubules, a property hitherto thought to have arisen as a VWF adaptation to the vasculature, is a feature of the amino-terminal region of mucin. This segment of the human intestinal gel-forming mucin (MUC2) was found to self-assemble into tubules with a striking resemblance to those of VWF itself. To facilitate a comparison, we determined the residue-resolution structure of tubules formed by the homologous segment of VWF. The structures of the MUC2 and VWF tubules revealed the flexible joints and the intermolecular interactions required for tubule formation. Steric constraints in full-length MUC2 suggest that linear filaments, a previously observed supramolecular assembly form, are more likely than tubules to be the physiological mucin storage intermediate. Nevertheless, MUC2 tubules indicate a possible evolutionary origin for VWF tubules and elucidate design principles present in mucins and VWF.


Investigation of Feedback Schedules on Speech Motor Learning in Older Adults.
The principles of motor learning (PML) emerged from studies of limb motor skills in healthy, young adults. The applicability of these principles to speech motor learning, and to older adults, is uncertain. The purpose of this study was to examine one PML, feedback frequency, and its effect on retention and generalization of a novel speech and comparable tracing task. Sixty older adults completed a speech motor learning task requiring the production of a novel phrase at speaking rates 2 times and 3 times slower than habitual rate. Participants also completed a limb motor learning task requiring the tracing of a sine wave 2x and 3x slower than habitual rate. Participants were randomly assigned to receive feedback every trial, every 5th trial, or every 10th trial. Mean absolute error was measured to examine immediate generalization, delayed generalization, and 2-day retention. Results suggested that feedback frequency did not have an effect on the retention and generalization of the speech or manual task, supporting the small but growing literature highlighting the constraints of generalizing the PML to other modalities and populations.


Frequency specificity of simultaneously recorded early and middle latency auditory evoked potentials.
Early and middle latency auditory evoked potentials were recorded simultaneously with a 2-channel wide-band recording technique. Latency and amplitude distributions of components V, Na and Pa were determined in 20 normal hearing adults for 3 different stimuli (click, plop, 500 Hz tone burst) presented at 70, 30 and 20 dB HL. Normative latency data of the middle latency components are presented. The detectability and amplitude of the Na-Pa complex were considerably larger than those of wave V at 20 and 30 dB HL for the two low frequency stimuli. Ten ears exhibiting high frequency sensorineural hearing loss were examined. In these cases, only click-evoked Na and Pa, but not wave V, were observed down to 20 dB HL. Also, the click latency-intensity function of Na-Pa was found to be similar to the normal low frequency latency functions. The results consistently suggested that the low frequency contributions to wave V cancel in phase because of short duration, whereas the duration of the Na-Pa complex allows for a positive superimposition of all frequency bands. Since these differences in frequency specificity of the EAEP and the MAEP affect latency, care is advised for the interpretation of inter-peak latencies.


Herpes management and prophylaxis.
In summary, clinical management of HSV infection depends on an individual having an appropriate diagnosis and receiving proper counseling, follow-up of both themselves and their sexual partners, as well as having an opportunity to discuss with more knowledgeable physicians, the options for antiviral management should their individual problems and sequelae of genital HSV infection require it. Although some of the aspects of management such as HSV type 2 serology testing or continuous antiviral therapy will be limited in their availability to individuals worldwide because of cost, it is possible to improve knowledge among physicians about diagnosis. Furthermore, it is necessary to ensure that facilities for patient information and counselling are available. These latter facilities are often provided extremely efficiently by self-help or voluntary groups so it is vital that physicians understand and ensure that they obtain details of such groups to pass on to their patients.


Human lung cDC1 drive increased perforin-mediated NK cytotoxicity in chronic obstructive pulmonary disease.
In chronic obstructive pulmonary disease (COPD), lung natural killer cells (NKs) lyse autologous lung epithelial cells in vitro, but underlying mechanisms and their relationship to epithelial cell apoptosis in vivo are undefined. Although this cytolytic capacity of lung NKs depends on priming by dendritic cells (DCs), whether priming correlates with DC maturation or is limited to a specific DC subset is also unknown. We recruited ever-smokers (≥10 pack-years; n = 96) undergoing clinically indicated lung resections. We analyzed lung NKs for cytotoxic molecule transcripts and for cytotoxicity, which we correlated with in situ detection of activated Caspase-3/7+ airway epithelial cells. To investigate DC priming, we measured lung DC expression of CCR2, CCR7, and CX3CR1 and cocultured peripheral blood NKs with autologous lung DCs, either matured using lipopolysaccharide (LPS) (nonobstructed smokers) or separated into conventional dendritic cell type-1 (cDC1) versus cDC type-2 (cDC2) (COPD). Lung NKs in COPD expressed more perforin (P < 0.02) and granzyme B (P < 0.03) transcripts; inhibiting perforin blocked in vitro killing by lung NKs. Cytotoxicity in vitro correlated significantly (Sr = 0.68, P = 0.0043) with numbers of apoptotic epithelial cells per airway. In nonobstructed smokers, LPS-induced maturation enhanced DC-mediated priming of blood NKs, reflected by greater epithelial cell death. Although CCR7 expression was greater in COPD in both cDC1 (P < 0.03) and cDC2 (P = 0.009), only lung cDC1 primed NK killing. Thus, rather than being intrinsic to those with COPD, NK priming is a capacity of human lung DCs that is inducible by recognition of bacterial (and possibly other) danger signals and restricted to the cDC1 subset.


Three-dimensional imaging of congenital disorders involving the orbit.
Three-dimensional imaging of the orbit and its adnexa provides an excellent topographic visualization of the deformity or tumor extent. This helps comprehension, communication, education, and documentation in the process of treating the patient. This article briefly describes the technique of three-dimensional imaging and classifies congenital orbital deformities which are extensively illustrated with relevant case material.


Antigen-dependent IL-12 signaling in CAR T cells promotes regional to systemic disease targeting.
Chimeric antigen receptor (CAR) T cell therapeutic responses are hampered by limited T cell trafficking, persistence, and durable anti-tumor activity in solid tumors. However, these challenges can be largely overcome by relatively unconstrained synthetic engineering strategies. Here, we describe CAR T cells targeting tumor-associated glycoprotein-72 (TAG72), utilizing the CD28 transmembrane domain upstream of the 4-1BB co-stimulatory domain as a driver of potent anti-tumor activity and IFNγ secretion. CAR T cell-mediated IFNγ production facilitated by IL-12 signaling is required for tumor cell killing, which is recapitulated by engineering an optimized membrane-bound IL-12 (mbIL12) molecule in CAR T cells. These T cells show improved antigen-dependent T cell proliferation and recursive tumor cell killing in vitro, with robust in vivo efficacy in human ovarian cancer xenograft models. Locoregional administration of mbIL12-engineered CAR T cells promotes durable anti-tumor responses against both regional and systemic disease in mice. Safety and efficacy of mbIL12-engineered CAR T cells is demonstrated using an immunocompetent mouse model, with beneficial effects on the immunosuppressive tumor microenvironment. Collectively, our study features a clinically-applicable strategy to improve the efficacy of locoregionally-delivered CAR T cells engineered with antigen-dependent immune-modulating cytokines in targeting regional and systemic disease.


Does direct-to-consumer advertising affect patients' choice of pain medications?
In the United States, direct-to-consumer advertising (DTCA) has grown rapidly to promote prescription medications, including analgesics. Few studies in the literature directly examine the association between DTCA and patients' choice of pain medications. This article discusses how DTCA affects such choice from a behavioral perspective, because DTCA-prompted behaviors are important indicators of DTCA's influence. After DTCA exposure, patients may request prescriptions, seek further medication information, and ask about advertised conditions. Patients who suffer from pain may seek more communication with their health care providers because they are cautious about the information quality of DTCA, mainly because of the recall of rofecoxib (Vioxx; Merck, Whitehouse Station, NJ). However, the availability and DTCA of over-the-counter analgesics complicate their treatment choice. Patients could use DTCA as a tool to launch health communication and make an informed treatment choice with the guidance of their health care providers.


Medical management of otitis externa.
Successful medical management of otitis externa requires attention to all of the following points: 1. Diagnose and treat systemic or underlying disease(s). 2. Carefully clean and examine the entire ear canal before starting therapy. This includes removal of foreign bodies, parasites, hair, and other obstructions. 3. Apply carefully chosen topical preparations based on gross and microscopic examination. 4. Educate clients as to the causes of the otitis externa and their role in treating the disease. 5. Schedule regular follow-up examinations until the disease is completely cured. 6. Recommend preventive procedures such as drying ears after swimming, corrective surgery, and so on. 7. Use systemic treatment when indicated by chronicity of disease or the owner's inability to treat the patient.


Fast exocytosis and endocytosis triggered by depolarisation in single adrenal chromaffin cells before rapid Ca2+ current run-down.
The kinetics of exocytosis and membrane retrieval (endocytosis) were examined in bovine chromaffin cells using membrane capacitance measurement during whole-cell recording. At early times after breakthrough to the whole-cell recording mode, depolarisation for 1 s resulted in a fast (600 vesicles per s) exocytotic response and efficient membrane retrieval with a time constant of 25 s. The ability to activate fast exocytosis and retrieval was lost during intracellular dialysis, with a time constant of 40 s. At later times, a slow exocytotic response could be elicited with no membrane retrieval following single depolarisations. The wash-out of the responses appeared to be due to a rapid loss of a portion of the Ca2+ current. Trains of depolarisation at late times after breakthrough could elicit a fast (time constant 4 s) retrieval. These data show that in addition to a previously studied slow Ca(2+)-independent retrieval mechanism, chromaffin cells also possess an efficient and rapid retrieval pathway coupled to exocytosis that can be activated following depolarisation. The fast endocytosis appears to have a higher threshold for activation than exocytosis, probably due to a higher Ca2+ requirement. Rapid membrane retrieval appears to occur via a clathrin-independent pathway in chromaffin cells.


Seminal fructose in normal and infertile men.
Fructose levels and fructolysis index in human semen were analysed to assess a correlation, if any, between the levels of this glycolysable sugar and sperm concentration. Semen was collected from normospermic men and men with azoospermia or oligospermia. Seminal fructose levels were elevated in men with obstructive azoospermia and in men who remained azoospermic following vasoepididy mostomy done to correct epididymal blockage. Men with sperm concentration of less than 20 million/ml pre-operatively or following vasoepididy mostomy, showed significantly high levels of fructose and lower fructolysis index. Fructose levels in normospermic infertile men, as well as in men with normal sperm counts (more than 20 million/ml), were similar to that in men of proven fertility.


High-Dose Versus Standard-Dose Intensity-Modulated Radiotherapy With Concurrent Paclitaxel Plus Carboplatin for Patients With Thoracic Esophageal Squamous Cell Carcinoma: A Randomized, Multicenter, Open-Label, Phase 3 Superiority Trial.
The standard dose (SD) of definitive concurrent chemoradiotherapy (dCRT) remains 50.4 Gy in patients with esophageal cancer; a higher dose, when applied with conventional radiation therapy techniques, increases toxicities without improving survival. We investigated whether a high dose of 59.4 Gy using intensity-modulated radiation therapy (IMRT) would improve survival without increasing toxicities. Patients with inoperable thoracic esophageal squamous cell carcinoma (SCC) referred for dCRT were randomly assigned (1:1) to high-dose (HD) IMRT (59.4 Gy) or SD IMRT (50.4 Gy). Chemotherapy consisted of 6 cycles of concurrent weekly paclitaxel and carboplatin and a maximum of 2 cycles of consolidation chemotherapy. Nutritional intervention was implemented for patients with malnutrition on the basis of nutritional screening. The primary endpoint was median overall survival (mOS). Analyses were by modified intention to treat. Between April 30, 2016, and April 30, 2019, 167 patients were enrolled at 9 participating centers in China. Seventy-one patients in the HD and 73 patients in the SD groups were included in the analysis; 86.8% of the patients completed radiation therapy and 70.1% received 5 or 6 cycles of concurrent chemotherapy. The median follow-up was 36.0 months. The mOS was 28.1 and 26.0 months in the HD and SD arms, respectively (P = .54). A total of 7 treatment-related deaths were observed. Grade 3 or worse treatment-related toxicities were observed in 62% and 68.5% of the patients in the HD and SD arms, respectively (P = .675). For patients with inoperable thoracic esophageal SCC, a dose of 59.4 Gy did not improve survival compared with the SD of dCRT using IMRT.


[Optimization of the extraction technology of total flavonoids from Mulgedium tataricum using response surface analysis].
Response surface analysis methodology (RSM) was used to optimize the extraction technology of total flavonoids from Mulgedium tataricum. Ethanol concentration, solvent-solid ratio, extraction temperature and extraction time were selected as influencing factors during extraction. The experiment mathematical model was arranged according to Box-Behnken central composite experiment design. The optimal extraction conditions were as following: ethanol concentration 63.78%, solvent-solid ratio 40: 1, extraction temperature 90 degrees C, and extraction time 1.07 h, the predicted value of extraction rate of total flavonoids was 14.04% under the optimum conditions, and the experimental value was 13.79%. The combination of Box-Behnken design and response surface analysis can well optimize the extraction technology of total flavonoids from Mulgedium tataricum.


The Association Between Puberty Timing and Body Mass Index in a Longitudinal Setting: The Contribution of Genetic Factors.
We analyzed the contribution of genetic factors on the association between puberty timing and body mass index (BMI) using longitudinal data and two approaches: (i) genetic twin design and (ii) polygenic scores (PGS) of obesity indices. Our data were derived from Finnish cohorts: 9080 twins had information on puberty timing and BMI and 2468 twins also had genetic data. Early puberty timing was moderately associated with higher BMI in childhood in both boys and girls; in adulthood these correlations were weaker and largely disappeared after adjusting for childhood BMI. The largest proportion of these correlations was attributable to genetic factors. The higher PGSs of BMI and waist circumference were associated with earlier timing of puberty in girls, whereas weaker associations were found in boys. Early puberty is not an independent risk factor for adult obesity but rather reflects the association between puberty timing and childhood BMI contributed by genetic predisposition.


Caffeic acid phenethyl ester preferentially enhanced radiosensitizing and increased oxidative stress in medulloblastoma cell line.
Caffeic acid phenethyl ester (CAPE), an active component of propolis, was recently reported to have radiosensitizing effects on medulloblastoma (MB) cells. However, the mechanisms of radiosensitivity involved in medulloblastoma cells are still unclear. The specific aim of this study was to investigate the role of CAPE-induced oxidative stress to influence of radiosensitivity and anti-proliferative effects in medulloblastoma cells. Medulloblastoma (Daoy) cells were treated with CAPE in different concentrations and assessed for cell viability. The following were also evaluated: migratory ability, reduced glutathione (GSH) level, reactive oxygen species (ROS) level, nuclear factor-kappaB (NF-kappaB) activity, and apoptosis in CAPE alone, radiation alone, or radiation combined with CAPE in Daoy cells. The results indicated that CAPE inhibited the growth of Daoy cells. CAPE treatment in Daoy cells could effectively decrease glutathione reductase and significantly increase glutathione peroxidase. Radiation-activated NF-kappaB was reversed by CAPE pretreatment. Finally, the result of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay showed that CAPE treatment can enhance radiation-induced apoptosis in Daoy cells. Our study demonstrated the anti-proliferative and radiosensitizing effects of CAPE on MB cells, which may be achievable through depleting GSH, increased ROS activity, and inhibiting NF-kappaB activity.


High-Anion-Gap Metabolic Acidosis During a Prolonged Hospitalization Following Perforated Diverticulitis: An Educational Case Report.
The metabolic acidoses are generally separated into 2 categories on the basis of an anion gap calculation: high-anion-gap and normal anion-gap metabolic acidosis. When a high-anion-gap metabolic acidosis (HAGMA) is not clearly explained by common etiologies and routine confirmatory testing, specialized testing can definitively establish rare diagnoses such as 5-oxoproline, d-lactate accumulation, or diethylene glycol toxicity. A 56-year-old woman had a prolonged hospital admission following perforated diverticulitis requiring sigmoid resection. Her hospitalization was complicated by feculent peritonitis and surgical wound dehiscence needing prolonged broad-spectrum antibiotics and wound debridements. She developed acute kidney injury and HAGMA in the hospital. Chart review showed that she received a large cumulative dose of acetaminophen during her hospital stay. Laboratory studies showed markedly increased serum 5-oxoproline causing HAGMA. Patient was admitted to the intensive care unit and treated with N-acetylcysteine and renal replacement therapy. After admission to the intensive care unit, the patient continued to require vasopressor and ventilatory support for septic shock and a ventilator-associated pneumonia. After an initial recovery and resolution of her HAGMA, she subsequently suffered recurrent aspirations which were fatal. 1. The acronym GOLD MARK is useful when assessing patients with HAGMA and most causes of HAGMA can be established with routine testing.2. When the etiology of HAGMA remains unclear, additional testing can be required to diagnose rare causes of HAGMA.3. Rare causes of HAGMA are diethylene glycol, 5-oxoproline, and d-lactate accumulation.4. Acidosis secondary to 5-oxoproline accumulation can occur even with "therapeutic" doses of acetaminophen in patients receiving it regularly for a prolonged period and who have depleted glutathione stores.5. Risk factors for glutathione depletion include malnutrition, older age, sepsis, pregnancy, multiple chronic illnesses, and chronic kidney disease.


Ways in which SWL affects oxidant/antioxidant balance.
Many damages can occur in the kidneys following extracorporeal shock wave lithotripsy (SWL) and some of these damages are thought to emerge as a result of oxidative stress. The objective of this study is to evaluate the impact of SWL application on kidneys in stone patients on oxidative stress parameters in 24-h urine. The study included 20 patients scheduled for SWL. total antioxidant capacity (TAC) and total oxidant status (TOS) measurements were made on 24-h urines of the patients before and after SWL. The oxidative stress index (OSI) was calculated based on these values. Following SWL, while TOS increased in 24-h urine (p = 0.006), TAC remained unchanged (p = 0.312). OSI increased following the procedure (p = 0.004). SWL application disrupts the oxidant/antioxidant balance in favor of oxidants, which leads to oxidative stress. TOS, TAC and OSI may be useful in the determination of the existence of oxidative stress following SWL.


Linear Solvation Energy Relationships as classifiers in non-target analysis--a capillary liquid chromatography approach.
For the identification of unknowns in environmental samples, Linear Solvation Energy Relationships (LSERs) are applied as classifiers for exclusion of candidate compounds with equal exact mass. By prediction of the Chromatographic Hydrophobicity Index (CHI) from retention time of the unknown compound on a gradient system using a C18 column and the description of interactions by LSER descriptors, an exclusion of isomers, with different retention behaviour and interaction terms is possible. The example of several isomers, with the empirical formula C₁₂H₁₀O₂, demonstrates the application as classifier and indicates limits of this prognosis.


Distinct retinoid metabolic functions for alcohol dehydrogenase genes Adh1 and Adh4 in protection against vitamin A toxicity or deficiency revealed in double null mutant mice.
The ability of class I alcohol dehydrogenase (ADH1) and class IV alcohol dehydrogenase (ADH4) to metabolize retinol to retinoic acid is supported by genetic studies in mice carrying Adh1 or Adh4 gene disruptions. To differentiate the physiological roles of ADH1 and ADH4 in retinoid metabolism we report here the generation of an Adh1/4 double null mutant mouse and its comparison to single null mutants. We demonstrate that loss of both ADH1 and ADH4 does not have additive effects, either for production of retinoic acid needed for development or for retinol turnover to minimize toxicity. During gestational vitamin A deficiency Adh4 and Adh1/4 mutants exhibit completely penetrant postnatal lethality by day 15 and day 24, respectively, while 60% of Adh1 mutants survive to adulthood similar to wild-type. Following administration of a 50-mg/kg dose of retinol to examine retinol turnover, Adh1 and Adh1/4 mutants exhibit similar 10-fold decreases in retinoic acid production, whereas Adh4 mutants have only a slight decrease. LD(50) studies indicate a large increase in acute retinol toxicity for Adh1 mutants, a small increase for Adh4 mutants, and an intermediate increase for Adh1/4 mutants. Chronic retinol supplementation during gestation resulted in 65% postnatal lethality in Adh1 mutants, whereas only approximately 5% for Adh1/4 and Adh4 mutants. These studies indicate that ADH1 provides considerable protection against vitamin A toxicity, whereas ADH4 promotes survival during vitamin A deficiency, thus demonstrating largely non-overlapping functions for these enzymes in retinoid metabolism.


Comparison of innate and Th1-type host immune responses in Oesophagostomum dentatum and Trichuris suis infections in pigs.
This study investigated the details of the innate and Th1/Treg-type-associated host immune responses in Trichuris suis and Oesophagostomum dentatum mono- and co-infected pigs and in vitro in stimulated porcine dendritic cell cultures. Forty-eight pigs were allocated into a 2-factorial design with two groups trickle-inoculated with 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (O). Another group (OT) was infected with both parasites. Group C remained uninfected. Expression of innate and Th1/Treg-cell-associated genes in gut mucosa and associated lymph nodes was determined by qPCR at necropsy day 35 and 71. Gene expression showed suppressed/inhibited Th1 and Treg-type immune reactions, in accordance with previous findings of a predominant Th2-type immune response to both nematodes. The in vitro part examined the production of TNF-α in porcine dendritic cells (DC) exposed to T. suis and/or O. dentatum excretory/secretory (E/S) products. Further, binding capacity and structure of E/S products were characterized. Glycan and lectin-binding capacity were generally lower in O. dentatum E/S products compared to T. suis which may explain the earlier found weaker Th2 response to the former. Surprisingly, O. dentatum E/S products induced a significant (P < 0·0001) increase in TNF-α DC production, potentially indicating a new mode of helminth-host immune response interaction.


Verbally Highlighting Extrinsic Causes of Novel Social Disparities Helps Children View Low-Status Groups as Structurally Disadvantaged Rather Than Personally Inferior.
As part of their "essentialist" intuitions, young children tend to form personal attributions for observed intergroup differences - attributing them to groups' intrinsic natures or inborn characteristics. Much research has linked this essentialist view of social groups with prejudiced attitudes. However, less research has explored children's capacity to form structural attributions for observed intergroup differences - attributing them to groups' extrinsic circumstances or access to opportunities - or how structural attributions relate to social attitudes. Structural attributions could enable children to view low-status groups as extrinsically disadvantaged rather than intrinsically inferior. We were interested in whether verbally highlighting the extrinsic causes of novel social status disparities could support young children in forming structural attributions for them, thereby mitigating the formation of prejudice toward novel low-status groups. To investigate, we introduced participants (n=106 5- and 6-year olds) to novel social status disparities that could be attributed to either intrinsic or extrinsic causes, and we framed the disparities in either intrinsic, neutral, or extrinsic terms. We then assessed children's attributions for the disparities (through two measures: explanations and interventions) and their social attitudes toward the groups (through two measures: friendship preferences and prize allocations). Results indicated that participants tended to provide mostly personal attributions overall but that extrinsic framing led them to provide significantly more structural attributions. Extrinsic framing did not significantly impact social attitudes overall, but exploratory analyses revealed that it impacted participants' friendship preferences in particular. Together, results suggest that verbally highlighting extrinsic causes can disrupt children's intuitive tendency toward personal attributions, with promising implications for their views of low-status groups.


Postsynaptic Potentiation in Mouse Motor Synapses Induced by ATP Accumulation in Synaptic Cleft.
In mouse motor synapses, a non-selective purinoceptor antagonist suramin increased the quantum content of endplate potentials (EPP) without changing the time course of synaptic potentials. An ectonucleotidase inhibitor ARL 67156 had no effect on the amplitude and quantum content of EPP and miniature endplate potentials (mEPP) evoked by single stimuli, but significantly prolonged their duration. Long-term high-frequency stimulation of the nerve in the presence of ARL 67156 persistently increased the amplitude and duration of EPP during the train of impulses, but did not change their quantum content. ATP-γ-S, a non-hydrolyzed ATP analogue, significantly increased the amplitudes and prolonged the rising and falling phases of EPP and mEPP. The ATP-induced postsynaptic potentiation in neuromuscular transmission can result from the increase in ATP content and its longer presence in the synaptic cleft.


Morphological but not functional changes of the carotid artery are associated with the extent of coronary artery disease in patients with preserved left ventricular function.
The atherosclerotic process is associated with both morphological and functional changes in the carotid artery. We evaluated the relationship between these parameters of the carotid artery and the extent of coronary artery disease (CAD) in patients with preserved left ventricular function. The study population consisted of 104 stable patients with CAD who had preserved left ventricular function (left ventricular ejection fraction >or=45%). All patients underwent carotid ultrasound for evaluation of carotid artery plaque score defined by the sum of plaque thickness, maximum percent area stenosis, and carotid arterial stiffness index beta calculated by a combination of changes in carotid arterial diameter and blood pressure. Plaque score and percent area stenosis correlated with the extent of CAD defined as the number of diseased coronary vessels (P<0.001 and 0.002, respectively), but arterial stiffness beta did not (P=0.39). Using logistic regression analyses adjusting for confounding coronary risk factors and arterial stiffness beta, plaque score and percent area stenosis were independently correlated with multivessel CAD (P=0.001 and 0.004, respectively). Carotid artery plaque burden, but not arterial stiffness, is associated with the extent of CAD, suggesting morphological rather than functional changes in the carotid artery may be a more accurate predictor of the extent of CAD and multivessel CAD independent of left ventricular function.


Long term outcome of anastomotic leakage in patients undergoing low anterior resection for rectal cancer.
The influence of anastomotic leakage (AL) on local recurrence rates and survival in rectal cancer remains controversial. The aim of this study was to analyze the effect of asymptomatic anastomotic leakage (AAL) and symptomatic anastomotic leakage (SAL) on short- and long-term outcome after curative rectal cancer resection. All patients who underwent surgical resection of non-metastatic rectal cancer with curative intent from January 2005 to December 2017 were retrospectively analyzed. Short-term morbidity, long-term functional and oncological outcomes were compared between patients with SAL, AAL and without AL (WAL). Overall, 200 patients were included and AL was observed in 39 (19.5%) patients (10 AAL and 29 SAL) with a median follow-up of 38.5 months. Rectal cancer location and preoperative neoadjuvant treatment was similar between the three groups. Postoperative 30-day mortality rate was nil. The permanent stoma rate was higher in patients with SAL or AAL compared to WAL patients (44.8 and 30% vs 9.3%, p < 0.001). The mean wexner continence grading scale was significantly different between AAL (11,4 ± 3,8), SAL (10,3 ± 0,6) and WAL (6,4 ± 4,7) groups (p = 0.049). The 3 and 5-year overall and disease-free survival rates were similar between the 3 groups (86.6% /84% vs 100%/100% vs 76%/70 and 82.9%/77% vs 100%/100% vs 94.7%/88.3% for patients with SAL, AAL, and WAL, p = 0.480 and p = 0.527). The permanent stoma rate was significant higher in patients with SAL or AAL compared to WAL patients. AL did not impair long-term oncological outcome.


Preliminary analysis of double-negative T, double-positive T, and natural killer T-like cells in B-cell chronic lymphocytic leukemia.
B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the expansion of CD5+ malignant B lymphocytes. Recent discoveries have shown that double-negative T (DNT) cells, double-positive T (DPT) cells, and natural killer T (NKT)-cells may be involved in tumor surveillance. A detailed immunophenotypic analysis of the peripheral blood T-cell compartment of 50 patients with B-CLL (classified in three prognostic groups) and 38 healthy donors (as controls) matched for age was performed. The samples were analyzed by flow cytometry using a stain-lyse-no wash technique and a comprehensive six-color antibody panels. Our data confirmed a reduction in percentage values and an increase in absolute values of T lymphocytes in patients with B-CLL, as already reported. In particular, DNT, DPT, and NKT-like percentages were significantly lower than in the controls, except for NKT-like in the low-risk prognostic group. Moreover, a significant rise in the absolute counts of DNT cells in each prognostic group and in the low-risk prognostic group of NKT-like cells was found. A significant correlation of the absolute values of NKT-like cells in the intermediate-risk prognostic group versus B cells was observed. Furthermore, we analyzed whether the increase in T cells was related to the subpopulations of interest. Only DNT cells were positively correlated with the increase in CD3+ T lymphocytes, regardless of the stage of the disease, supporting the hypothesis that this T-cell subset plays a key role in the immune T response in B-CLL. These early results supported that DNT, DPT, and NKT-like subsets may be related to disease progression and should encourage further studies aimed at identifying the potential immune surveillance role of these minority T subpopulations.


The Conjugation of Antibodies for the Simultaneous Detection of Surface Proteins and Transcriptome Analysis at a Single-Cell Level.
Transcriptome analysis at a single-cell level with single-cell RNA sequencing (scRNA-seq) is a powerful method for detailed characterization of heterogeneous cell populations. Recent developments have enabled parallel analysis of both transcript and protein levels by using antibodies conjugated to barcoded oligonucleotides. These antibodies enable protein levels to be converted into nucleotide format, allowing the sequencing-based detection of both modalities at single-cell level. Here we present a simple and reliable method for conjugation of oligonucleotides with antibodies and a protocol for their use in single-cell transcriptome sequencing.


Different susceptibility of whole casein components to enzymatic phosphorylation by two forms of rat liver 'casein kinase'.
The phosphorylation of the single casein subfractions occurring when whole casein is incubated with [gamma-32P]ATP in the presence of two different rat liver 'casein kinases', both cyclic AMP-insensitive, has been studied. "Casein kinase TS", active on both threonine and serine residues of whole casein, was found to be active towards a minor protein fraction, running slightly ahead of beta-casein during gel electrophoresis, and accounting for most, if not all, of the [32P]Thr residues labeled in whole casein ("[32P]Thr-rich fraction"). The [32P]Ser residues labeled by this enzyme were recovered in an heterogeneous "[32P]Ser-rich fraction" including alphas1-casein together with minor alphas fractions, following alphas1-casein during gel electrophoresis. "Casein kinase S", on the other hand, active only towards serine residues of whole casein, is active almost exclusively towards the minor alphas casein fractions, with the exclusion of both the "[32P]Thr-rich fraction" and alphas1-casein itself. Therefore, of the major casein components, beta- and K-caseins apparently play a quite unimportant role in the overall phosphorylation of whole casein by both the protein kinases tested, while alphas1-casein itself, unlabeled by casein kinase S, accounts for no more than 20--30% of 32P incorporated in the presence of casein kinase TS.


Containment measures limit environmental effects on COVID-19 early outbreak dynamics.
Environmental factors are well known to affect spatio-temporal patterns of infectious disease outbreaks, but whether the rapid spread of COVID-19 across the globe is related to local environmental conditions is highly debated. We assessed the impact of environmental factors (temperature, humidity and air pollution) on the global patterns of COVID-19 early outbreak dynamics during January-May 2020, controlling for several key socio-economic factors and airport connections. We showed that during the earliest phase of the global outbreak (January-March), COVID-19 growth rates were non-linearly related to climate, with fastest spread in regions with a mean temperature of ca. 5 °C, and in the most polluted regions. However, environmental effects faded almost completely when considering later outbreaks, in keeping with the progressive enforcement of containment actions. Accordingly, COVID-19 growth rates consistently decreased with stringent containment actions during both early and late outbreaks. Our findings indicate that environmental drivers may have played a role in explaining the early variation among regions in disease spread. With limited policy interventions, seasonal patterns of disease spread might emerge, with temperate regions of both hemispheres being most at risk of severe outbreaks during colder months. Nevertheless, containment measures play a much stronger role and overwhelm impacts of environmental variation, highlighting the key role for policy interventions in curbing COVID-19 diffusion within a given region. If the disease will become seasonal in the next years, information on environmental drivers of COVID-19 can be integrated with epidemiological models to inform forecasting of future outbreak risks and improve management plans.


[Thyroid hormone analogs: an important biological supply and new therapeutic possibilities].
Thyroid hormones [predominantly 3, 5, 3 -I- iodothyronine (T3)] regulate cholesterol and lipoprotein metabolism but cardiac effects restrict their use as hypolipidemic drugs. New molecules have been developped which target specifically the thyroid hormone receptor ss, predominant isoform in liver. The first thyroid hormone agonist, called GC1, has selective actions compared to T3. In animals, GC1 reduced serum cholesterol and serum triglycerides, probably by stimulation important steps in reverse cholesterol transport. Other selective thyromimetic, KB- 2115 and KB - 141 have similar effects. Another class of thyroid hormone analogs, the thyronamines have emerged recently but the basic biology of this new class of endogenous thyroid hormone remains to better understood. Therefore, these molecules may be a potentially treatment for obesity and reduction cholesterol, triglycerides and lipoprotein (a). To date the studies in human are preliminary. Tolerance and efficacy of these drugs are still under investigation.


Recent trends in cancer survival across Europe between 2000 and 2004: a model-based period analysis from 12 cancer registries.
Monitoring population-based cancer survival is an essential component in the evaluation of cancer control, but subject to an inherent delay in the reporting of the most recent survival estimates with traditional techniques of analysis. We examined survival trends between the years 2000 and 2004 for 20 common cancers based on follow-up data from 12 cancer registries from diverse areas of Europe using model-based period analysis techniques. Between 2000 and 2004, marked rises were seen in 5-year relative survival amongst patients with prostate, breast and colorectal cancer, which were statistically significant in 10, 8 and 7 of the 12 participating cancer registries, respectively. For cancer sites amenable to effective early detection and treatment, major geographical differences in patient prognosis still persisted, with a lower survival generally observed in Eastern European countries. Model-based period analysis enables the timely monitoring of recent trends in population-based cancer survival. For colorectal and breast cancers, the identified rises in survival are probably (at least partly) explained by the improvements in clinical care and the management of the disease. Nevertheless, persisting geographic differences do point to the potential for a further reduction in the burden of cancer throughout Europe, towards which improvements in diverse areas of care, including secondary prevention, access to advances in treatment as well as subspecialisation and regionalisation of oncologic care may all contribute.


Accelerated intimal hyperplasia in aortocoronary internal mammary vein grafts in minipigs.
More than 50% of aortocoronary saphenous vein grafts are occluded 10 years after surgery. Intimal hyperplasia is the initial critical step in the progression toward occlusion. Internal mammary veins, which are physiologically prone to less hydrostatic pressure, may undergo an accelerated progression to intimal hyperplasia and thus be suitable for investigation of the mechanisms of aortocoronary vein graft disease. Six minipigs underwent aortocoronary bypass grafting using standard cardiopulmonary bypass and cardioplegic arrest. Mammary vein were grafted in a reversed manner from ascending aorta to left anterior descending coronary artery (LAD). The proximal LAD was ligated, rendering the anterior left ventricle vein graft-dependent. Minipigs were killed after 4 weeks, and vein grafts were harvested. Histological and immunohistological investigation were performed with respect to morphometric analysis, endothelial damage/dysfunction (v-Willebrand-factor (vWF)), smooth muscle cells (alpha-smooth actin) and proliferation rate (proliferation marker Ki 67). Mean intimal area of vein grafts was increased compared to ungrafted mammary veins. Intimal hyperplasia in vein grafts was characterized by massive accumulation of smooth muscle cells with a high proliferation rate and endothelial perturbation. Significant (p = 0.001) intimal hyperplasia of the grafted mammary vein compared to the ungrafted mammary vein was found. These changes were absent in ungrafted mammary veins. The present study demonstrates a pig model of aortocoronary vein graft intimal hyperplasia which is characterized by an accelerated progression within internal mammary veins. The model is suitable to investigate the pathophysiology of aortocoronary vein graft intimal hyperplasia as well as therapeutic approaches.


Genetic evidence that the causal association of educational attainment with reduced risk of Alzheimer's disease is driven by intelligence.
Alzheimer's disease (AD) is predicted to affect 132 million people by 2050. Targeting modifiable lifestyle risk factors that are associated with an increased risk of AD could prevent a large proportion of dementia cases, allowing people to reach the end of their life dementia free. However, evidence obtained from the observational studies does not take into account how risk factors are correlated with one another, and whether they causally contribute to increased AD risk. In this study, we determine whether the relationship between previously speculated AD risk factors and AD susceptibility is consistent with causality using large-scale genetic data. We focus on educational attainment (EA), intelligence and household income which have been previously shown to be causally associated with AD. Using GWAS-by-subtraction and Multivariable Mendelian Randomization we show that of these, only the cognitive component of EA (intelligence) is independently causally associated with AD. This work has ramifications for the modifiability of lifestyle risk factors for AD.


Effect of gastric secretagogues on the formation of inositol phosphates in isolated gastric cells of the rat.
The effects of compounds affecting gastric acid secretion were studied on the formation of inositol phosphates after prelabelling with [3H]-inositol in enriched gastric parietal cells of the rat, prepared by isopycnic centrifugation with Percoll. In cell preparations with 60 to 70% parietal cells, carbachol (10(-6)-10(-2) M) enhanced the accumulation of [3H]-inositol monophosphate ([3H]-IP1), [3H]-inositol bisphosphate ([3H]-IP2) and [3H]-inositol trisphosphate ([3H]-IP3) in a concentration-dependent manner, an effect which was antagonized by 10(-8) M atropine. Li+ (0.5-30 mM) enhanced the basal and carbachol-induced accumulation of all three [3H]-inositol phosphates, the formation of [3H]-IP1 being more sensitive to Li+ than those of [3H]-IP2 and [3H]-IP3. The concentration of Ca2+ in the incubation medium did not affect the relative stimulation of the accumulation of [3H]-inositol phosphates by carbachol, although the basal formation was higher in the presence of Ca2+ in the medium. In the absence of added Ca2+, the incorporation of [3H]-inositol into phospholipids was increased--an effect which was further enhanced by the addition of EGTA to the medium. Gastrin and pentagastrin (10(-8)-10(-5) M) enhanced the formation of [3H]-inositol phosphates, although they were clearly less effective than carbachol. Histamine (10(-6)-10(-3) M) had no effect of its own, but slightly attenuated the effect of carbachol. Cholecystokinin octapeptide (10(-9)-10(-6) M) slightly increased the formation of [3H]-inositol phosphates. Indomethacin (10(-4) M) had no consistent effect on the basal and carbachol-induced accumulation of [3H]-inositol phosphates, nor did prostaglandin E2 (10(-5) M) modify it. Adrenaline (10(-3) M), 5-hydroxytryptamine (10(-3) M), forskolin (10(-5) M), vasopressin (10(-5) M), angiotensin II (10(-5) M) and bombesin (10(-9)-10(-6) M) were all without effect. We suggest that the hydrolysis of inositol phospholipids may be involved in the signal transduction mechanism by which the activation of the muscarinic and gastrin receptors on the parietal cells leads to Ca2+ mobilization and the stimulation of hydrogen ion secretion.


Axonal function in a family with episodic ataxia type 2 due to a novel mutation.
Episodic ataxia type 2(EA-2) is a rare, autosomal dominant disorder characterised by recurrent episodes of ataxia and dysarthria,due to mutations in the CACNA1A gene on chromosome 19 encoding voltage-dependent Ca2+ channels. The aim of the present study was to explore whether axonal membrane properties, assessed using nerve excitability techniques, were abnormal in patients with EA-2 . Nerve excitability techniques were applied to the median nerve of three individuals from three generations of a single family, all of whom had typical features of EA-2. This family was found to have a novel mutation at codon 1451 of the Ca2+ channel alpha 1A subunit. Nerve excitability testing demonstrated significant abnormalities,with all patients outside the normal 95 % confidence limits in having a high rheobase and reduced early hyperpolarizing threshold electrotonus. On average there were also significant reductions in refractoriness,late sub excitability and early depolarizing threshold electrotonus. Mathematical modelling indicated that a similar pattern of abnormalities may result from a reduced voltage dependence of slow K+ channels (KCNQ channels). There are significant and distinctive changes in peripheral nerve excitability in EA-2 patients,which are presumably induced indirectly. These findings raise the possibility that excitability testing may prove a convenient screening test for patients with this suspected channelopathy.


Executive Summary: Guidelines for the Prevention and Treatment of Opportunistic Infections in HIV-Exposed and HIV-Infected Children: Recommendations From the National Institutes of Health, the Centers for Disease Control and Prevention, the HIV Medicine Association of the Infectious Diseases Society of America, the Pediatric Infectious Diseases Society, and the American Academy of Pediatrics.
The Guidelines for the Prevention and Treatment of Opportunistic Infections (OIs) in HIV-Exposed and HIV-Infected Children in the United States were developed by a panel of specialists in pediatric HIV infection and infectious diseases from the U.S. government and academic institutions, intended for use by clinicians and health care workers providing medical care for HIV-exposed and HIV-infected children in the United States. For each OI, pediatric specialists with subject matter expertise reviewed the literature for new information since the last guidelines were published (2009) and then proposed revised recommendations that were reviewed and approved by the full Panel and endorsing governmental agencies and professional organizations. This executive summary highlights the most important, rated recommendations for each OI from the full Guidelines document.


Vascular Disease Burden, Outcomes and Benefits with Empagliflozin in Heart Failure: Insights From the EMPEROR-Reduced Trial.
The presence of ischemic heart disease impacts prognosis in patients affected by heart failure and reduced ejection fraction (HFrEF). It is not well known how the extent of vascular disease impacts prognoses and responses to therapy in this setting. In this post hoc analysis of the EMPEROR-Reduced trial, outcomes and the effects of empagliflozin, were assessed in study participants according to the extent (none vs mono1 vs poly [≥ 2] vascular bed) of vascular disease. Vascular disease was defined as investigator-reported coronary artery disease (CAD), peripheral artery disease (PAD) and cerebrovascular disease at baseline. Cox proportional-hazards models were used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs). Incidence rates are presented per 100 person-years (py) of follow-up. Of the 3730 study participants enrolled, 1324 (35.5%) had no vascular disease, 1879 (50.4%) had monovascular disease, and 527 (14.1%) had polyvascular disease. Participants with polyvascular disease tended to be older and male and to have had histories of hypertension, diabetes and smoking. In the placebo arm, a significantly higher risk for cardiovascular death existed in those with polyvascular disease (HR 1.57, 95% CI1.02, 2.44, compared to those with no vascular disease). In adjusted analysis, the benefit of empagliflozin in cardiovascular death or hospitalization due to HF, HF hospitalization, cardiovascular death, renal composite endpoint, estimated glomerular filtration slope changes, and health status scores were seen across the 3 groups (interaction P > 0.05 for all) but were attenuated in those with polyvascular disease. Adverse events were higher in those with polyvascular disease, but no major differences were noted between empagliflozin or placebo assignment in the 3 groups. In patients with HFrEF, the extent of vascular disease is associated with the risk for adverse cardiovascular outcomes. Empagliflozin offers cardiovascular and renal benefits in HFrEF across the extent of vascular disease, but this benefit is attenuated in those with polyvascular disease.


ZHX2 inhibits proliferation and promotes apoptosis of human lung cancer cells through targeting p38MAPK pathway.
To investigate the effect of ZHX2 on lung cancer cells proliferation and apoptosis. The mRNA and protein expression of ZHX2 were detected by qRT-PCR and western blot, respectively. The human lung cancer cells were divided into Control, NC, ZHX2, SB, and ZHX2 + Ani groups. The cell proliferation was detected by CCK-8 assay and the cell migration and invasion were detected by Transwell assay. Cell apoptosis was detected by flow cytometry. Apoptosis and p38MAPK signaling pathway related proteins were detected by western blot. The nude mice model of lung cancer xenograft was constructed. The tumor volume and tumor weight were measured. The expression of PCNA protein in tumor tissues was detected by immunohistochemistry. The apoptosis of tumor cells was detected by TUNEL staining. The ZHX2 and p38MAPK signaling pathway related proteins in tumor tissues were detected by western blot. The expression of ZHX2 gene and protein in the cancer cell lines were significantly decreased. Compared with control and NC groups, the cells proliferation, migration and invasion were inhibited in ZHX2 and SB groups, while the apoptosis and apoptosis related proteins were increased (p< 0.05). Meanwhile, compared with ZHX2 group, the tumor growth rate, volume, weight, the percentage of PCNA-positive cells, and p-P38 MAPK/P38 MAPK were increased significantly in ZHX2 + Ani group, while the apoptotic index and the expression of MMP-9 protein were significantly decreased (p< 0.05). ZHX2 could inhibit proliferation and promote apoptosis of lung cancer cells by inhibiting p38MAPK signaling pathway.


Public versus private: evidence on health insurance selection.
This paper models health insurance choice in Chile (public versus private) as a dynamic, stochastic process, where individuals consider premiums, expected out-of pocket costs, personal characteristics and preferences. Insurance amenities and restrictions against pre-existing conditions among private insurers introduce asymmetry to the model. We confirm that the public system services a less healthy and wealthy population (adverse selection for public insurance). Simulation of choices over time predicts a slight crowding out of private insurance only for the most pessimistic scenario in terms of population aging and the evolution of education. Eliminating the restrictions on pre-existing conditions would slightly ameliorate the level (but not the trend) of the disproportionate accumulation of less healthy individuals in the public insurance program over time.


Divalent cations induce protofibril gelation.
Soluble fibrin oligomers (protofibrils) undergo phase change merely by adding 1-2 mM Ca2+ or 25-100 microM Zn2+. The cation-induced "protofibrin" clots appear similar to normally formed fibrin gels. Maximal clot turbidity of protofibrin gels increases with cations in a concentration-dependent manner. Magnesium (less than 0.5 mM) is ineffective in inducing protofibril gelation. Turbidity and degree of polymerization (DP) [indirectly expressed as AT (activation time)] appear to be positively correlated, regardless of whether the divalent cation is Ca2+ or Zn2+. Cross sections of Ca2+-induced protofibrin fibers are approximately 6-18-fibrin-monomers-thick. With both Ca2+ and 40 microM Zn2+, fiber cross section increases to 30-50 monomers thick. Negatively stained Zn2+-and Ca2+-induced protofibrin gels exhibit banding periodicity of approximately 240 A, similar to that of normally generated fibrin gels. Regions of lateral merging of individual segments of the protofibrin leads to increased cross section of the fiber and forms a branch required for gelation. These findings indicate that the rate of coagulation and the ultimate thickness of the fibers both relate to lateral associative processes of protofibrils, which are augmented by physiologic concentrations of 2+ and Zn2+.


The Design and Evaluation of Clinical Pathway for Disease Management to Maximize Public Health Benefit.
As an implementation strategy, the actual functional scope of clinical pathway (CPW) can be mastered is the basis for evaluating and improving CPW management, which is very important for bridging the evidence-to-practice gap. Here, we propose and discuss a design and evaluation model for CPWs through some theoretical lenses that may help the designer master the actual functional scope of CPWs and accumulate and refine effective key management steps of a specific disease. We can thus determine the best-optimized design with the largest net benefit that allows definition of the core management steps for a specific disease.


Chemical composition, mineral profile, and functional properties of Canna (Canna edulis) and Arrowroot (Maranta spp.) starches.
The aim of the present study was to evaluate some chemical and mineral characteristics and functional and rheological properties of Canna and Arrowroot starches produced in the Venezuelan Andes. Canna starch showed a higher (P < 0.05) moisture, ash, and crude protein content than arrowroot starch, while crude fiber, crude fat, and amylose content of this starch were higher (P < 0.05). Starches of both rhizomes own phosphorus, sodium, potassium, magnesium, iron, calcium, and zinc in their composition. Phosphorus, sodium, and potassium are the higher in both starches. Water absorption, swelling power, and solubility values revealed weak bonding forces in Canna starch granules; this explained the lower gelatinization temperature and the substantial viscosity development of Canna starch during heating. Arrowroot starch showed a higher gelatinization temperature measure by DSC, than Canna starch and exhibited a lower value of DeltaH. Both starches show negative syneresis. The apparent viscosity of Canna starch was higher (P < 0.05) than the Arrowroot starch values. The size (wide and large) of Canna starch granules was higher than arrowroot starch. From the previous results, it can be concluded that Canna and Arrowroot starches could become interesting alternatives for food developers, depending on their characteristics and functional properties.


Identification of a major GTP-binding protein in bovine aortic smooth muscle cytosol as the rhoA gene product.
In bovine aortic smooth muscle, about 50% of total GTP-binding activity was present in the cytosol fraction. A major GTP-binding protein (G protein) with a Mr value of about 21,000 (21K G) in this fraction was purified to near homogeneity and characterized. 21K G bound maximally about 0.8 mol of [35S]guanosine 5'-(3-O-thio)triphosphate/mol of protein with a Kd value of about 20 nM. 21K G showed GTPase activity with a turnover number of about 0.007 min-1. 21K G was ADP-ribosylated by botulinum ADP-ribosyltransferase and about 0.4 mol of ADP-ribose was maximally incorporated into 1 mol of 21K G. 21K G and the bovine brain rhoA gene product (rhoA p21) were eluted at the same retention time on C4 reversed-phase high performance liquid chromatography and migrated at the same positions on two-dimensional gel electrophoresis. These results indicate that the major G protein in bovine aortic smooth muscle cytosol is rhoA p21.


Ru(bpy)32+ as a photoinduced oxidase mimic for colorimetric detection of biothiols.
We have found that tris (2,2'-bipyridyl) ruthenium (II) (Ru(bpy)32+) possesses a high photo-induced oxidase-like activity and is capable of catalyzing the color reaction of 3,3',5,5'-tetramethylbenzidine (TMB) with dissolved oxygen. Ru(bpy)32+ has a catalytic constant (Kcat) that is twice as high as that of fluorescein, 170 and 275-fold higher than that of 9-mesityl-10-methyl acridine and Eosin Y, respectively. Electron spin resonance spectroscopy (ESR) and radical scavenging experiments have verified the major active radicals involved in the color reaction are •OH. A colorimetric biothiol assay has been successfully developed for the oxidase-like activity of Ru(bpy)32+ can be suppressed by sulfhydryl compounds. A linear dependence between the decrease in absorbance and the logarithm of thiol concentrations can be found ranging from 5.0 to 50 μM, with a detection limit of 1.0 μM. This work reveals a new oxidase mimic with high catalytic activity and will facilitate the utilization of this oxidase mimic in biochemical analysis.


Chelant-aided enhancement of lead mobilization in residential soils.
Chelation of metals is an important factor in enhancing solubility and hence, availability to plants to promote phytoremediation. We compared the effects of two chelants, namely, ethylenediaminetetraacetic acid (EDTA) and ethylenediaminedisuccinic acid (EDDS) in enhancing mobilized lead (Pb) in Pb-based paint contaminated residential soils collected from San Antonio, Texas and Baltimore, Maryland. Batch incubation studies were performed to investigate the effectiveness of the two chelants in enhancing mobilized Pb, at various concentrations and treatment durations. Over a period of 1 month, the mobilized Pb pool in the San Antonio study soils increased from 52 mg kg(-1) to 287 and 114 mg kg(-1) in the presence of 15 mM kg(-1) EDTA and EDDS, respectively. Stepwise linear regression analysis demonstrated that pH and organic matter content significantly affected the mobilized Pb fraction. The regression models explained a large percentage, from 83 to 99%, of the total variation in mobilized Pb concentrations.


Validation of reference genes for qPCR studies on Caco-2 cell differentiation.
Validation of reference gene expression stabilities is a prerequisite for reliable normalization of qPCR data. The present study assessed the variation of six reference genes (ACTB, GAPDH, B2M, HPRT1, SDHA, YWHAZ) in Caco-2 cells under the influence of different growth supports and cultivation periods. Genes were ranked according to their stability using the geNorm software. To verify the influence of reference gene selection, ALPI gene expression during differentiation was quantified using the most or the least stable reference gene for normalization. Experimental conditions significantly affected the expression levels of reference genes. Whereas GAPDH and ACTB were revealed as most stable genes, SDHA was the least stable one. The extent of ALPI gene expression was significantly changed by the selection of the reference gene. This study provides a basis for qPCR studies related to both the differentiation process of Caco-2 cells and the elucidation of cell behaviour influenced by surface modifications.


USP28 and USP25 are downregulated by Vismodegib in vitro and in colorectal cancer cell lines.
Deubiquitinase USP28 plays a crucial role in tumorigenesis by enhancing the stabilities of multiple cancer-related proteins including c-Myc, Notch1, and LSD1, and has become an attractive target for anticancer drug development. However, to date, only a few of USP28-targeted active compounds have been developed, and the active compound-binding pocket in USP28 has not been experimentally revealed yet. In this study, bioassay-based high-throughput screening was applied to discover USP28-targeted inhibitors from the commercially available drug library. Vismodegib, an inhibitor of Hedgehog signaling pathway and FDA-approved drug for the treatment of basal cell carcinoma, was found to exhibit inhibition activity against USP28 (IC50 : 4.41 ± 1.08 μm). Multiple biophysical and biochemical techniques including NMR, ITC, thermal shift assay, HDX-MS, and site-directed mutagenesis analysis were then used to characterize the interaction between Vismodegib and USP28. The binding pocket in USP28 for Vismodegib, which is mainly composed of two helical structures spanning D255-N278 and N286-Y293, was revealed. According to the possible binding pose generated by HDX-MS data-defined molecular docking, the binding cavity occupied by Vismodegib in USP28 aligns well with one of the reported-binding pockets in USP7 for its inhibitors. Furthermore, cellular assays were conducted to confirm that Vismodegib could interact with the evolutionarily related deubiquitinases USP28 and USP25 and downregulate the levels of the two enzymes' substrate proteins c-Myc, Notch1, and Tankyrase-1/2.


M2 Macrophage-Derived Exosomal lncRNA AFAP1-AS1 and MicroRNA-26a Affect Cell Migration and Metastasis in Esophageal Cancer.
Exosomes from cancer cells or immune cells, carrying bio-macromolecules or long non-coding RNAs (lncRNAs), participate in tumor pathogenesis and progression by modulating the microenvironment. This study aims to explore the function of M2 macrophage-derived exosomes on the invasion and metastasis of esophageal cancer (EC) with the involvement of the lncRNA AFAP1-AS1/microRNA-26a (miR-26a)/activating transcription factor 2 (ATF2) axis. We found that lncRNA AFAP1-AS1 could specifically bind to miR-26a, thus affecting the expression of miR-26a, and ATF2 was the direct target of miR-26a. Compared with M1 macrophage-derived exosomes, M2 macrophage-derived exosomes exhibited higher AFAP1-AS1 and ATF2 expression and lower miR-26a expression. Moreover, extracellular AFAP1-AS1 could be moved to KYSE410 cells via being incorporated into M2 macrophage-derived exosomes. M2 macrophage-derived exosomes could downregulate miR-26a and promote the expression of ATF2 through high expression of AFAP1-AS1, thus promoting the migration, invasion, and lung metastasis of EC cells; M2-exosomes upregulating AFAP1-AS1 or downregulating miR-26a ameliorated this effect. In summary, M2 macrophage-derived exosomes transferred lncRNA AFAP1-AS1 to downregulate miR-26a and upregulate ATF2, thus promoting the invasion and metastasis of EC. Targeting M2 macrophages and the lncRNA AFAP1-AS1/miR-26a/ATF2 signaling axis represents a potential therapeutic strategy for EC.


Induction and regulation of interleukin-6 gene expression in rat astrocytes.
Cells that produce interleukin-6 (IL-6) require the presence of signaling molecules since this cytokine is not normally constitutively expressed. It is now established that astrocytes produce IL-6; however, the precise inducing molecules and the kinetics of their action have not yet been clearly identified. In the current study, we show that either interleukin-1 beta (IL-1 beta) or tumor necrosis factor-alpha (TNF-alpha) exert a strong inducing signal for IL-6 in primary rat astrocytes. When the two cytokines are added together the response is synergistic, suggesting that each cytokine may induce IL-6 gene expression by different pathways. Interferon-gamma (IFN-gamma) does not affect IL-6 expression although if it is added in conjunction with IL-1 beta, an augmented induction of IL-6 occurs. In addition to the cytokines, bacterial lipopolysaccharide (LPS) and the calcium ionophore, A23187, induce IL-6 expression. IL-6 expression can be blocked by the glucocorticoid analogue, dexamethasone. IL-6 induction by LPS/Ca2+ ionophore is more sensitive to the suppressive effects of dexamethasone than is IL-6 induction by TNF-alpha/IL-1 beta. Cycloheximide (CHX), an inhibitor of protein synthesis, markedly increased levels of IL-6 mRNA in both unstimulated and stimulated astrocytes, indicating that ongoing protein synthesis is not required for astrocyte IL-6 gene expression. We propose that astrocyte-produced IL-6 may have a role in augmenting intracerebral immune responses in neurological diseases such as multiple sclerosis (MS), AIDS dementia complex (ADC), and viral infections. These diseases are characterized by infiltration of lymphoid and mononuclear cells into the central nervous system (CNS), and intrathecal production of immunoglobulins. IL-6 may act to promote terminal differentiation of B cells in the CNS, leading to immunoglobulin synthesis.


High-dose-rate brachytherapy alone post-hysterectomy for endometrial cancer.
To evaluate the outcome of post-hysterectomy adjuvant vaginal high-dose-rate (HDR) brachytherapy. A retrospective analysis was performed on a series of 143 patients with endometrial cancer treated with HDR brachytherapy alone post-hysterectomy from 1985 to June 1993. Of these patients, 141 received 34 Gy in four fractions prescribed to the vaginal mucosa in a 2-week period. The median follow-up was 6.9 years. Patients were analyzed for treatment parameters, survival, local recurrence, distant relapse, and toxicity. Five-year relapse free survival and overall survival was 100% and 88% for Stage 1A, 98% and 94% for Stage IB, 100% and 86% for Stage IC, and 92% and 92% for Stage IIA. The overall vaginal recurrence rate was 1.4%. The overall late-toxicity rate was low, and no RTOG grade 3, 4, or 5 complications were recorded. These results are similar to reported international series that have used either low-dose-rate or HDR brachytherapy. The biological effective dose was low for both acute and late responding tissues compared with some of the HDR brachytherapy series, and supports using this lower dose and possibly decreasing late side-effects with no apparent increased risk of vaginal recurrence.


Quality of life in patients with established inflammatory bowel disease: a UK general practice survey.
The current understanding of quality of life impairment in inflammatory bowel disease has largely been derived from selected populations and may not reflect the experience of patients in the community, where fewer than half are likely to be under specialist care. To describe the health-related quality of life in a community-based sample of patients with established inflammatory bowel disease and explore its association with the type and extent of disease, gender, age, material deprivation and other factors. Adults with established inflammatory bowel disease were identified systematically from the records of 23 family practices in north-east England. The health-related quality of life was assessed by self-completion of the UK Inflammatory Bowel Disease Questionnaire. Five hundred and fifty-six patients were sent the questionnaire and 409 (74%) gave usable replies. Lower scores (worse quality of life) were significantly associated with female gender, Crohn's disease, more extensive disease (ulcerative colitis) and being under specialist care. The mean health-related quality of life score was significantly lower in patients resident in more deprived districts, independent of the type and extent of disease. Most patients with established inflammatory bowel disease showed only minor impairment of their health-related quality of life. On average, women and those with Crohn's disease were relatively more affected. Clinicians responsible for the care of patients with inflammatory bowel disease should be aware of these more vulnerable groups.


Chip-based digital Polymerase Chain Reaction as quantitative technique for the detection of PIK3CA mutations in breast cancer patients.
PIK3CA is a gene frequently mutated in breast cancer. With the FDA approval of alpelisib, the evaluation of PIK3CA for activating mutations is becoming routinely. Novel platforms for gene analysis as digital PCR (dPCR) are emerging as a potential replacement for the traditional Sanger sequencing. However, there are still few studies on chip-based dPCR to detect mutations in tumor samples. Thus, this cross-sectional study aimed to assess the sensibility of a chip-based dPCR to detect and quantify PIK3CA mutations and compare its performance with Sanger sequencing. Tumor samples from 57 breast cancer patients (22 pre-treatment samples, 32 tumors after neoadjuvant chemotherapy, and three lymph nodes) were collected and analyzed by Sanger sequencing and dPCR for the three PIK3CA most relevant mutations (p.E545K, p. H1047R, and p. H1047L). Digital PCR sensitivity, specificity, and overall performance were estimated by contingency tables, receptor operator characteristic (ROC), and area under the curve (AUC). Association of PIK3CA mutations with clinicopathological variables was conducted. Sanger sequencing identified PIK3CA mutations in six patients (10.5%), two with p. H1047R, and four with p. E545K. Digital PCR confirmed those mutations and identified 19 additional patients with at least one mutation. Comparison between dPCR and Sanger sequencing showed a sensitivity of 100% (95% CI 53-100%), and a specificity of 84.2% (95% CI 83-84.2%). Besides, p. H1047R mutation detected by dPCR showed a significant association with breast cancer phenotype (p = 0.019) and lymphatic nodes infiltration (p = 0.046). Digital PCR showed a high sensitivity to detect mutations in tumor samples and it might be capable to detect low-rate mutations and tumor subpopulations not detected by Sanger sequencing.


Characterization of estrogen receptors and associated protein kinase activity by high-performance hydrophobic-interaction chromatography.
We have determined that high-performance hydrophobic-interaction chromatography (HPHIC) with weakly hydrophobic columns permit the rapid separation of the labile isoforms of estrogen receptor proteins. Previously we reported the use of the SynChrom propyl 500 column for HPHIC of steroid receptors. However, due to the strongly hydrophobic characteristics of the ligand, [125I]iodoestradiol-17 beta, and the receptor protein, organic solvent was required in the mobile phase for greater recovery of receptor proteins. Here, we report separation of steroid receptors from human breast tumors and rat uteri, using the Beckman CAA-HIC, a non-ionic polyether-bonded column, without the need for organic solvents and with virtually 100% recoveries. Receptors were extracted in 10 mM phosphate buffer (pH 7.4). Maximum resolution and separation were achieved when a descending salt gradient of ammonium sulfate in phosphate buffer (pH 7.4) was used (2-0 M in 30 min). Estrogen receptor (ER) was resolved into two isoforms with tR = 22 +/- 1 min (n = 16, designated as peak I) and 27.5 +/- 0.5 min (n = 14, designated as peak II) and a purification of five- to twenty-fold in a single pass. Free steroid was eluted at tR = 35 +/- 1 min (n = 4). Separation was dependent on adjusting the ionic strength of cytosol to 1.5 M ammonium sulfate. ER, purified by HPHIC, retained ligand binding capacity and exhibited protein kinase activity, which was dominant in the less hydrophobic peak I (tR = 22 min) when immunoprecipitated with the monoclonal antibody D547. This method of rapidly purifying ER with high retention of biological activity may now be applied to the study of the molecular interrelationships of steroid receptor isoforms.


[Influence of the "distance to thrombus" in acute middle cerebral artery occlusion : A predictor for the clinical outcome after endovascular treatment].
The therapy strategy of patients with acute stroke of the middle cerebral artery (MCA) is influenced by the location of the occlusion. The purpose of this study was to analyze the clinical outcome in patients with acute ischemic MCA occlusion according to the location of occlusion who underwent endovascular treatment (EVT). A total of 54 patients (age 73 ± 15 years; 59% female) with acute ACM occlusion treated with EVT were included. In coronary reformatted CT angiography images, the distance to the thrombus (DT), i.e. the distance from the carotid T to the beginning of the thrombus, was measured. Correlations between DT, clinical symptoms, and clinical outcome of patients who underwent EVT were analyzed. DT correlated with clinical symptoms measured by the National Institutes of Health Stroke Scale (NIHSS; p = 0.017; R = -0.324) at baseline. DT also correlated with the modified Rankin scale after 90 days (90-day mRS; p = 0.014; R = -0.333). DT was a predictor for a good clinical outcome (mRS after 90 days) after EVT; odds ratio 1.113 (P = 0.02; 95% confidence interval [CI] 1.017-1.219). A DT >10 mm correlated significantly (p = 0.036) with a good clinical outcome (90-day mRS ≤2). DT correlates with the clinical symptoms of patients with acute MCA occlusion. In addition, DT is an independent predictor of the clinical outcome of patients suffering from acute stroke due to MCA occlusion.


Release mechanism of omega-3 fatty acid in κ-carrageenan/polydextrose undergoing glass transition.
A high-solid matrix of κ-carrageenan with polydextrose was developed to entrap α-linolenic acid, which is an omega-3 bioactive compound. Physicochemical analysis of this system utilised modulated DSC, dynamic oscillation in shear, ESEM, FTIR and WAX diffraction. The carbohydrate matrix was conditioned through an extensive temperature range to induce changes in molecular morphology and identify the network glass transition temperature. Thermally induced variation in phase morphology was employed to rationalise transportation patterns of the bioactive compound within the high-solid preparation. Thus, experimental observations using UV-vis spectroscopy modelled diffusion kinetics to document the mobility arresting effect of the vitrifying matrix on the micro-constituent. Within the glass transition region, results argue that free volume theory is the molecular process governing structural relaxation. Further, Less Fickian diffusion follows well the rate of molecular transport of α-linolenic acid as a function of time and temperature of observation in the condensed matrix.


Characterization of HCT116 human colon cancer cells in an orthotopic model.
Colorectal cancer metastases result in a significant number of cancer related deaths. The molecular mechanisms underlying this complex, multi-step pathway are yet to be completely elucidated. In the absence of any transgenic models of colon cancer metastases, an in vivo model system that fulfills the rate limiting steps of metastasis (local invasion and distant colony formation) is needed. The purpose of this study was to characterize the behavior of a human colon cancer cell line, HCT116 in an orthotopic model. HCT116 cells were transfected with green fluorescence protein and subcutaneously injected into BALB/c nude male mice. Once xenografts were established, they were excised and orthotopically implanted into 32 other male BALB/c nude mice using microsurgical techniques. Animals were serially imaged and euthanized at 6-8 weeks post-implantation. Tissues were procured and processed for hematoxylin and eosin analysis. All 32 animals demonstrated primary tumor growth, invasion and peritoneal spread. Liver metastases were identified in 15/32 (47%), and lung metastases were confirmed in 13/32 (41%). In total, 19/32 (59%) animals demonstrated distant metastatic colony formation. This orthotopic model of colon cancer fulfills the rate limiting steps of local invasion and distant colony formation in the process of metastases. HCT116 human colon cancer cell line in this in vivo model system provides a tool to dissect the molecular mechanism involved in the metastatic cascade.


The politics of prostate cancer screening.
The controversial recent recommendation by the United States Preventive Services Task Force (USPSTF) against prostate-specific antigen (PSA) screening for early-stage prostate cancer has caused much debate. Whereas USPSTF recommendations against routine screening mammography in younger women resulted in fierce public outcry and eventual alteration in the language of the recommendation, the same public and political response has not been seen with PSA screening for prostate cancer. It is of paramount importance to ensure improved efficiency and transparency of the USPSTF recommendation process, and resolution of concerns with the current USPSTF recommendation against PSA screening for all ages.


Modern role of clinical ultrasound in liver abscess and echinococcosis.
In the Western world, liver abscesses are predominantly of pyogenic origin and can present a wide range of ultrasonographic features, from a solid mass to a cystic cavity. Amoebic abscesses are endemic in tropical areas and typically round single lesions. Echinococcal cyst is diagnosed by ultrasound (US). Serology is particularly useful when pathognomonic US features are absent. Treatment is determined based on the WHO ultrasonographic classification, and puncture, aspiration, injection, re-aspiration (PAIR) is a US-guided therapeutic option. Hepatic alveolar echinococcosis presents on US as a pseudotumoral mass frequently with calcifications that may invade the biliary tree and portal and hepatic veins.


The 4Cs of adaptation tracking: consistency, comparability, comprehensiveness, coherency.
Adaptation tracking seeks to characterize, monitor, and compare general trends in climate change adaptation over time and across nations. Recognized as essential for evaluating adaptation progress, there have been few attempts to develop systematic approaches for tracking adaptation. This is reflected in polarized opinions, contradictory findings, and lack of understanding on the state of adaptation globally. In this paper, we outline key methodological considerations necessary for adaptation tracking research to produce systematic, rigorous, comparable, and usable insights that can capture the current state of adaptation globally, provide the basis for characterizing and evaluating adaptations taking place, facilitate examination of what conditions explain differences in adaptation action across jurisdictions, and can underpin the monitoring of change in adaptation over time. Specifically, we argue that approaches to adaptation tracking need to (i) utilize a consistent and operational conceptualization of adaptation, (ii) focus on comparable units of analysis, (iii) use and develop comprehensive datasets on adaptation action, and (iv) be coherent with our understanding of what constitutes real adaptation. Collectively, these form the 4Cs of adaptation tracking (consistency, comparability, comprehensiveness, and coherency).


The identification of 3-methylcholanthrene-9,10-dihydrodiol as an intermediate in the binding of 3-methylcholanthrene to DNA in cells in culture.
Two metabolites of 3-methylcholanthrene (3MC), previously shown to be precursors of the DNA-bound form of 3MC observed in embryo cells in culture, were prepared from 3MC by microsomal metabolism and isolated by high pressure liquid chromatography (HPLC). From HPLC analysis of the metabolites of 3MC, from mass spectrometric analysis and from comparison with the fluorescence spectra of all 5 possible dihydrodiols of the alkylated benzanthracenes, it was deduced that one of the precursor metabolites was a 9,10-dihydrodiol of 3MC while the other was a 1 or 2-hydroxy derivative thereof.


Circ-CCS is identified as a cancer-promoting circRNA in lung cancer partly by regulating the miR-383/E2F7 axis.
Increasing biomolecules have been found to be involved in the lung cancer development. This study will perform the function and mechanism analyses of a novel circular RNA copper chaperone for superoxide dismutase (circ-CCS) in lung cancer. Circ-CCS, microRNA-383 (miR-383) and E2F transcription factor 7 (E2F7) were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was detected using Cell Counting Kit-8 (CCK-8). Clonal ability was measured by colony formation assay. Cell apoptosis was determined via flow cytometry. Cell migration and invasion were assessed by transwell assay. Detection of protein was completed using western blot. Xenograft assay was used for the functional analysis of circ-CCS in vivo. The binding between targets was proved by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. E2F7 protein level was also examined by Immunohistochemistry (IHC) analysis in human tissues. Circ-CCS was upregulated in lung cancer and could predict poor prognosis. Downregulation of circ-CCS inhibited lung cancer cell growth and metastasis while promoted apoptosis in vitro, and suppressed tumorigenesis of lung cancer in vivo. Circ-CCS had sponge effect on miR-383 and the function of si-circ-CCS was achieved by upregulating miR-383. E2F7 was a target gene of miR-383 and its downregulation was responsible for the anti-cancerous role of miR-383 in lung cancer. Circ-CCS could elevate E2F7 expression via interacting with miR-383. Circ-CCS was shown to facilitate lung cancer progression via the miR-383/E2F7 axis, exhibiting the pivotal value of circ-CCS in diagnosis and treatment of lung cancer.


In-home use of a hybrid closed loop achieves time-in-range targets in preschoolers and school children: Results from a randomized, controlled, crossover trial.
To obtain additional information on the incremental differences between using a sensor-augmented pump (SAP) without automated insulin delivery (AID), using it with predictive low-glucose management (PLGM) or as hybrid closed loop (HCL), in preschool and school children. We conducted a monocentric, randomized, controlled, two-phase crossover study in 38 children aged 2-6 and 7-14 years. The primary endpoint was the percentage of time in range (TIR) of 70-180 mg/dl. Other continuous glucose sensor metrics, HbA1c, patient-related outcomes (DISABKIDS questionnaire, Fear of Hypoglycaemia Survey) and safety events were also assessed. Results from 2 weeks of SAP, 8 weeks of PLGM and 8 weeks of HCL were compared using a paired t-test or Wilcoxon signed-rank test. Overall, we found a high rate of TIR target (>70%) achievement with HCL in preschool (88%) and school children (50%), with average times in Auto Mode of 93% and 87%, respectively. Preschool children achieved a mean TIR of 73% ± 6% (+8% vs. SAP, +6% vs. PLGM) and school children 69% ± 8% (+15% vs. SAP and + 14% vs. PLGM). Overall, HbA1c improved from 7.4% ± 0.9% to 6.9% ± 0.5% (P = .0002). Diabetes burden and worries and fear of hypoglycaemia remained at low levels, without significant changes versus PLGM. No events of severe hypoglycaemia or diabetic ketoacidosis occurred. Preschool children profit from AID at least as much as those aged 7 years and older. To ensure safe use and prescribing modalities, regulatory approval is also required for young children.