S100 as an immunohistochemically-detected marker with prognostic significance in endometrial carcinoma.
Several studies have indicated that dendritic cells (DC) participate in anti-tumor immunity, possibly influencing the course of malignant disease. We tested whether tumor infiltration by S100+ DC could be a prognostic marker for endometrial cancer. A retrospective study analyzing 115 tissue samples from patients with endometrial carcinoma and known histological grading as well as hormone receptor, Ki-67, Her-2/neu and p53 expression. Sections of paraffin-embedded endometrial tissue were immunohistochemically-stained with anti S100 antibody. Tumor infiltrating S100+ DC were counted via microscopic examination and calculated as S100+ DC per mm2 of tissue. Samples were divided into group 1: less than 10 S100+DC/mm2 (n = 44) and group 2: 10 or more S100+ DC/mm2 (n = 71). Correlation with clinico-pathological markers was calculated by Chi-square test. Compared to group 1, the DC-rich group 2 showed a higher level of differentiation (p=0.045), a lower overexpression of p53 (p=0.021) and less proliferation (p=0.028). DC infiltration was not correlated with Her-2/neu, hormone receptor status and FIGO-stage. Although no significant correlation could be seen, the DC-poor group samples seemed to correlate with a higher FIGO-stage compared to the DC-rich group. In uni- and multivariate analysis, DC infiltration proved to be a significant prognostic marker for adjusted survival but not for overall survival. Our results indicate that the immunohistochemical determination of S100+ DC could contribute to the identification of a high-risk subgroup and, therefore, would be a favorable prognostic factor for endometrial carcinoma. Our observation that pronounced DC infiltration is associated with good prognosis points to an important role of the host's immune system response for the clinical course of endometrial cancer.


The stabilizing and unstabilizing influences of neurogenic and vascular activities of the heart as related to sudden cardiac death.
Despite the frequency with which sudden cardiac death occurs in affluent societies, little is known about the precise mechanisms by which it is caused. Practically nothing is known about why sudden cardiac death occurs in one but not another person. It is difficult to escape the conclusion that in many instances, the final events occur almost by chance. Therefore, it would appear that every effort should be made by the physician to assist in preventing the changes, that is, cardiomegaly and myocardial ischemia, that appear to be potential markers of sudden cardiac death in many persons. Attention should be directed toward identifying and facilitating stabilizing neurogenic and vascular activities and identifying, controlling and reversing unstabilizing influences likely to foster sudden cardiac death. In the absence of a more complete knowledge of precise mechanisms, the efforts of the physician should be directed toward behavior modifications that appear to reduce the clinical manifestations of coronary heart disease which contribute to the development of cardiomegaly and myocardial ischemia.


[Establishment of human immortalized oral epithelial cell line HIO615 induced by HPV16 E6 and E7].
To establish an immortalized oral epithelial cell line. Normal human oral epithelial cells were transfected with HPV16E6/E7 open reading frames using recombinant retroviral system pLXSN. Expression of HPV16E6 and E7 protein were tested by Western blot in three kinds of cells. To define cellular biological characterization of HPV16E6/E7 transfected cells, a series analysis were performed, including protraction of growth curve, HE staining, immunocytochemical staining and scanning electron microscope observation. The tumorigenicity was assessed by colony formation and transplanting the cells into nude mice. Human oral epithelial cells transfected with HPVE6/E7 has been in culture for over 18 months. The cell line was named HIO615. Western blot analysis showed HIO615 expressed HPV16 E6 and E7 protein. HIOC were positive for cytokeratin, tonofibril and desmosome as observed by scanning electron microscope. The number of large colonies of dense multilayer cells was low (0.77%). No tumor developed in nude mice injected subcutaneously with HIOEC. A human immortalized oral epithelial cell line induced by HPV16E6 and E7 has been successfully established.


Nucleotide sequence and regulation of a human 90-kDa heat shock protein gene.
In order to define the mechanisms responsible for the regulated expression of a human 90-kDa heat shock protein (HSP90) gene, we have determined the complete genomic sequence of the gene encoding HSP90 beta, including 1102 base pairs upstream of the transcription initiation site. This gene consists of 12 exons and 11 introns. The exons range in size from 99 to 396 base pairs and the introns range from 91 to 1433 base pairs. Analysis of the 5' promoter region revealed TATA and CAAT consensus sequences. Additionally, Sp1 factor binding sites and presumptive heat shock element consensus sequences were found surrounding the transcription initiation site. Three out of the six identified potential heat shock elements reside within the first intron. The ability of the promoter region to respond to heat shock was examined by constructing a plasmid containing the 5' end of the HSP90 beta gene driving a neomycin phosphotransferase reporter gene. This plasmid was used to stably transform human K562 erythroleukemia cells. With heat shock, the abundance of HSP-neo transcripts increased, demonstrating that the promoter region of this gene contains the cis-acting DNA elements required for the heat shock response.


[Evaluation of pre- and post-operational esophageal manometric findings with achalasia].
To evaluate the manometric feature of pre- and post-operation with achalasia and the result of surgical myotomy through abdominal incision. The manometric studies were performed in 23 pre- and 11 post-operational patients, including 11 patients who received both pre- and post-operational studies. 24 hour pH monitoring was performed in 11 after surgical myotomy. Achalasia, a motor disorder of the esophageal body, and hypertension of the resting LES impaired relaxation. The incidence of reflux was 22.2%. Surgical myotomy through abdominal incision is an effective procedure. It is necessary to avoid LES incompetence or reflux esophagitis.


Stromal Vascular Fraction and Mesenchymal Stem Cells from Human Adipose Tissue: A Comparison of Immune Modulation and Angiogenic Potential.
In recent years, both stromal vascular fraction (SVF) from adipose tissue and mesenchymal stem cells (MSC) from adipose tissues were extensively used in both preclinical and clinical treatment for various diseases. Some studies reported differences in treatment efficacy between SVFs and MSCs in animals as well as in humans. Therefore, this study is aimed to evaluate the immune modulation and angiogenic potential of SVFs and MSCs from the same SVF samples to support an explanation when SVFs or MSCs should be used. The adipose tissue samples from ten female donors with consent forms were collected. SVFs from these samples were isolated according to the published protocols. The existence of mesenchymal cells that positive with CD44, CD73, CD90, and CD105 and endothelial progenitor cells that positive with CD31 and CD34 was determined using flow cytometry. Three samples of SVFs with similar percentages of mesenchymal cell portion and endothelial progenitor cell portion were used to isolate MSCs. Obtained MSCs were confirmed as MSCs using the ISCT minimal criteria. To compare the immune modulation of SVF and MSCs, the mixed lymphocyte assay was used. The lymphocyte proliferation, as well as IFN-gamma and TNF-alpha concentrations, were determined. To compare the angiogenic potential, the angiogenesis in quail embryo assay was used. The angiogenesis efficacy was measured based on the vessel areas formed in the embryos after 7 days. The results showed that all SVF samples contained the portions of mesenchymal cells and endothelial progenitor cells. MSCs from SVFs meet all minimal criteria of MSCs that suggested by ISCT. MSCs from SVFs efficiently suppressed the immune cell proliferation compared to the SVFs, especially at ratios of 1:4 (1 MSCs: 4 immune cells). MSCs also inhibited the IFN-gamma and TNF-alpha production more efficiently than SVFs (p < 0.05). However, in quail embryo models, SVFs triggered the angiogenesis and neovessel formation better than MSCs with more significant vessel areas after 7 days (p < 0.05). This study suggested that SVFs and MSCs have different potentials for immune modulation and angiogenesis. SVFs help the angiogenesis better than MSCs, while MSCs displayed the more significant immune modulation. These results can guide the usage of SVFs or MSCs in disease treatment.


Activity and distribution of protein kinase C in liver during the acute-phase response.
Activity and subcellular distribution of protein kinase C were estimated in liver cytosol and membrane fractions of rats carrying a turpentine-induced inflammation. Protein kinase C activity increases significantly 8 h after treatment in the membrane fraction, with concurrent reduction in the cytosol; 10 h after treatment the membrane-associated activity returns to normal, without concomitant recovery of that detected in the cytosol. The specific binding of phorbol dibutyrate to the liver membrane fraction increases but overall the effect is less evident and delayed in time. The changes are associated to alterations in the phosphorylation pattern of some liver proteins. Liver protein kinase C activity and intracellular distribution seem to be affected by a treatment which is known to induce an acute-phase response in the liver cells.


Coexistence of Ferroelectricity and Ferromagnetism in Fullerene-Based One-Dimensional Chains.
One-dimensional (1D) magnetoelectric multiferroics are promising multifunctional materials for miniaturized sensors, actuators, and memories. However, 1D materials with both ferroelectricity and ferromagnetism are quite rare. Herein, using first-principles calculations, a series of fullerene-based 1D chains, namely U2 C@C80 -M (M = Cr, Mn, Mo, and Ru) 1D chains with both ferroelectric (FE) and ferromagnetic (FM) properties is designed. Compared to individual U2 C@Ih (7)-C80 , the spontaneous polarization (Ps) in 1D chains is enhanced by about two to four times owing to the interaction between U2 C@Ih (7)-C80 fullerene and M (M = Cr, Mn, Mo, and Ru) atoms. Meanwhile, the introduction of transition metal atoms dopes electrons into U's 5f orbitals, leading to numerous intriguing magnetic properties, such as U2 C@C80 -Cr and U2 C@C80 -Mo as 1D ferromagnetic semiconductors, U2 C@C80 -Ru as 1D ferrimagnetic (FiM) semiconductor, and U2 C@C80 -Mn as 1D antiferromagnetic (AFM) semiconductor. Excitingly, it is found that magnetic ordering and electrical polarization can be modulated independently by linking different transition metal atoms. These findings not only broaden the range of 1D multiferroic materials, but also provide promising candidates for novel electronic and spintronic applications.


Mitochondrial HSP70 cognate-mediated differential expression of JNK1/2 in the pollution stressed grey mullets, Mugil cephalus.
Cells have evolved different networks of cellular stress responses to adapt during environmental changes and survive combating wide variety of stress. Mitochondrial heat shock protein 70 (mtHSP70) are essential for maintaining cellular homeostasis. c-Jun-N terminal kinase (JNK), a subfamily of MAPK, plays a key role in the transmission of extracellular signal for survival and death. In the present investigation, the differential expression of phosphorylated JNK1/2 with corresponding increases in heat shock transcription factor-1 (HSF-1) under high levels of B-cell lymphoma 2 (Bcl-2), HSP70 transcripts (twofold) and mtHSP70 has been correlated with fish survival in polluted Ennore estuary. This study suggests that in Ennore sample, the amplification in HSP70 transcripts along with corresponding mtHSP70 overexpression might be due to activation of JNK1/2 and HSF-1. This would influence the expression of Bcl-2 protein involved in mitochondrial survival under pollution-stressed condition.


Ecdysteroid levels during embryogenesis in the shrimp, Palaemon serratus (Crustacea Decapoda): quantitative and qualitative changes.
During the embryogenesis of Palaemon serratus profound changes in ecdysteroid concentrations were found. Ecdysteroid concentrations increase at the appearance of the Y-organ and a slight decrease is observed just before hatching. Two peaks of ecdysteroid concentration occur between these two events if Palaemon is reared at 19 degrees, but only one broad maximum is observed after rearing at 11-12 degrees. The pattern of ecdysteroids changes during embryogenesis; the ecdysone level decreases, while the 20-OH-ecdysone concentration and the amount of high polarity products increase. During the embryonic stage C the ratio of 20-OH-ecdysone to ecdysone was shown by HPLC analysis to steadily increase. After enzymatic hydrolysis of high-polarity products the main RIA-positive material elutes from reversed-phase HPLC columns as 20-OH-ecdysone.


Postnatal serotonin type 2 receptor blockade prevents the emergence of anxiety behavior, dysregulated stress-induced immediate early gene responses, and specific transcriptional changes that arise following early life stress.
Early life adverse experience contributes to an enhanced vulnerability for adult psychopathology. Recent evidence indicates that serotonin type 2 (5-HT(2)) receptor function, implicated in the pathophysiology of mood and anxiety disorders, is significantly enhanced in the maternal separation model of early life stress. We examined whether postnatal 5-HT(2) receptor blockade would prevent the consequences of maternal separation on anxiety behavior and dysregulated gene expression. Control and maternally separated litters received treatment with the 5-HT(2) receptor antagonist, ketanserin, or vehicle during postnatal life and were examined for effects on adult anxiety behavior, adult stress-induced immediate early gene expression responses, and transcriptional changes within the prefrontal cortex during postnatal life and in adulthood. Treatment with ketanserin during postnatal life blocked the long-lasting effects of maternal separation on anxiety behavior in the open field test and the elevated plus maze. Further, the dysregulated adult stress-induced expression pattern of the immediate early gene, Arc, observed in maternally separated animals was also prevented by postnatal ketanserin treatment. Ketanserin treatment normalized the alterations in the expression of specific genes in the prefrontal cortex of maternally separated animals, including changes in serotonin type 2A receptor messenger RNA expression during postnatal life and in genes associated with G-protein signaling in adulthood. Postnatal treatment with the 5-HT(2) receptor antagonist, ketanserin, blocked specific consequences of maternal separation, including anxiety behavior and dysregulated gene expression in the prefrontal cortex. Our results suggest that enhanced 5-HT(2) receptor function may contribute to the emergence of anxiety behavior and perturbed stress responses following early life stress.


Low Cardiovascular Disease Awareness in Chilean Women: Insights from the ESCI Project.
Although cardiovascular disease (CVD) is the leading cause of mortality in Latin American women, limited data exist on CVD perceptions in this population. This study aimed to assess CVD awareness and knowledge of women from Santiago, Chile. This was a cross-sectional study conducted in women 35 to 70 years old. A multistage probability sampling (stratified by age and socioeconomic level) was used for participant selection. Participants completed a home survey about knowledge of CVD, risk factors, and perceived risk (based on standardized questions from the American Heart Association awareness survey). 723 women participated in the study (mean age: 51 ± 9 years; 17.6% with high education level). Only 9.3% of the respondents mentioned CVD as women's primary health problem, whereas 22.7% and 16.1%, respectively, listed breast cancer and other cancers. When asked to identify the leading cause of women's death, only 14.4% identified CVD compared to 69.1% who recorded cancer. Older women (≥ 55 years) more likely identified CVD as the main cause of death: (OR 2.9: 95% CI = 1.8-4.5) versus younger women (<55 years). CVD family history was also associated with higher awareness of CVD as the leading cause of death (OR 1.7: 95% IC; p = 1.1-2.6). Instead, women with middle education level were less likely to mention CVD as the main women's killer. Chilean women from Santiago have a low awareness of CVD as the leading cause of death and do not recognize CVD as their prominent health problem. Efforts should focus on increasing awareness and knowledge about CVD especially in young women.


Thermal ecology of allopatric lizards (Sphenomorphus) in southeast Australia : II. Physiological aspects of thermoregulation.
Physiological aspects of thermoregulation are examined in each of four species which constitute the Sphenomorphus quoyi species complex (an allopatric group of lizards, with similar Voluntary temperatures, restricted to southeast Australia).The smallest species (S. kosciuskoi) has a restricted and disjunct distribution at high altitudes. The largest species (S. quoyi) is restricted to lower altitudes. The other two species (S. tympanum - warm temperate, S. tympanum-cool temperate are intermediate in size and their combined allopatric distribution extends from low to high altitudes.Percentage integument reflectance of S. kosciuskoi is slightly lower than for the values of the other species. The species with the highest percentage integument reflectance is S. quoyi.In controlled laboratory conditions, the rate of body temperature increase at 29.5 and 10.5° C is greater than the rate of body temperature decrease in all species. Rate of heating and rate of cooling is similar in all species.It is concluded that interspecific differences in these physiological aspects of thermoregulation are slight and the differential control of heat flux by physiological means cannot account for the maintenance of similar Voluntary temperatures, in these lizards, where different habitats are utilized.


Warming Amplification of Minimum and Maximum Temperatures over High-Elevation Regions across the Globe.
An analysis of the annual mean temperature (TMEAN) (1961-2010) has revealed that warming amplification (altitudinal amplification and regional amplification) is a common feature of major high-elevation regions across the globe against the background of global warming since the mid-20th century. In this study, the authors further examine whether this holds for annual mean minimum temperature (TMIN) and annual mean maximum temperature (TMAX) (1961-2010) on a global scale. The extraction method of warming component of altitude, and the paired region comparison method were used in this study. Results show that a significant altitudinal amplification trend in TMIN (TMAX) is detected in all (four) of the six high-elevation regions tested, and the average magnitude of altitudinal amplification trend for TMIN (TMAX) [0.306±0.086 °C km-1(0.154±0.213 °C km-1)] is substantially larger (smaller) than TMEAN (0.230±0.073 °C km-1) during the period 1961-2010. For the five paired high- and low-elevation regions available, regional amplification is detected in the four high-elevation regions for TMIN and TMAX (respectively or as a whole). Qualitatively, highly (largely) consistent results are observed for TMIN (TMAX) compared with those for TMEAN.


Growing dendrites enhance a neuron's computational power and memory capacity.
Neocortical pyramidal neurons have many dendrites, and such dendrites are capable of, in isolation of one-another, generating a neuronal spike. It is also now understood that there is a large amount of dendritic growth during the first years of a humans life, arguably a period of prodigious learning. These observations inspire the construction of a local, stochastic algorithm based on an earlier stochastic, homeostatic, Hebbian developmental theory. Here we investigate the neurocomputational advantages and limits on this novel algorithm that combines dendritogenesis with supervised adaptive synaptogenesis. Neurons created with this algorithm have enhanced memory capacity, can avoid catastrophic interference (forgetting), and have the ability to unmix mixture distributions. In particular, individual dendrites develop within each class, in an unsupervised manner, to become feature-clusters that correspond to the mixing elements of class-conditional mixture distribution. Error-free classification is demonstrated with input perturbations up to 40%. Although discriminative problems are used to understand the capabilities of the stochastic algorithm and the neuronal connectivity it produces, the algorithm is in the generative class, it thus seems ideal for decisions that require generalization, i.e., extrapolation beyond previous learning.


The estimation of pyrophosphate in urine with uridine-5'-diphosphoglucose pyrophosphorylase.
The concentration of pyrophosphate in urine has been determined using the enzyme uridine-5'-diphosphoglucose pyrophosphorylase. The technique is relatively quick and easy to perform. Reproducibility of results was good, and results from recovery experiments were excellent. The mean concentration of pyrophosphate found in early morning urine samples from 14 normal adults was identical with that reported by other workers using an ion-exchange method to measure pyrophosphate. The pyrophosphate concentration was related approximately to the phosphate concentration.


Direct inhibitory effects of estrogens on rat Leydig cells in vitro.
In dispersed rat interstitial cells in vitro both natural and synthetic estrogens inhibited the action of pituitary luteinizing hormone (LH), as assessed by testosterone production. The estrogens also inhibited dibutyryl cyclic AMP induced steroidogenesis, suggesting that one point of inhibition could be distal to the formation of cyclic AMP in the cells. Diethyl stilbestrol and its clinically used sodium phosphate derivative (Honvol), also affected hormone-receptor interaction when tested with rat testicular homogenates. Among estradiol, estradiol benzoate, Honvol and diethyl stilbestrol only the latter at high concentration had toxic effects on Leydig cells as noted from loss of thier viability.


Photoinduced interaction between fluorescein ester derivatives and CdS colloid.
The photoinduced interaction of fluorescein ester derivatives, fluorescein's butyl ester (FL4) and fluorescein's anthraquinone-methyl ester (FL-AQ), and colloidal CdS was examined by absorption, fluorescence spectroscopy, and photoinduced ESR spectroscopy. It is found that FL4 and FL-AQ molecules are adsorbed on the surfaces of CdS colloid by an electrostatic interaction forming the surface complex of the type CdS-FL4 or CdS-FL-AQ. The apparent association constant (K(app)) and the degree of association (alpha) of CdS-FL4 obtained from absorption spectra are 2.25 x 10(4) M(-1) and 0.78, respectively. The values of K(app) and alpha of CdS-FL4 as determined from fluorescence spectra are 1.54 x 10(4) M(-1) and 0.82, respectively, which matches well with that determined from the absorption spectra changes. And the values of K(app) and alpha of CdS-FL-AQ obtained from absorption spectra are 4.18 x 10(4) M(-1) and 0.83, respectively. These data indicate that there is a strong interaction between the dye and the CdS particle surface. But there was no evidence for interfacial electron transfer from FL4 or FL-AQ to colloidal CdS by photoinduced ESR experiments. The fluorescence quenching is due to the formation of a nonfluorescent complex. The related phenomena are discussed in this paper.


Measurement of spermidine/spermine-N1-acetyltransferase activity by high-performance liquid chromatography with N1-dansylnorspermine as the substrate.
We developed a nonradioactive assay to measure spermidine/spermine N(1)-acetyltransferase (SSAT) activity by high-performance liquid chromatography (HPLC). N(1)-dansylnorspermine was prepared and evaluated as a substrate of acetylation with acetyl-CoA by SSAT in rat hepatoma (HTC) cells. Kinetic studies revealed that the K(m) values of N(1)-dansylnorspermine and acetyl-CoA were approximately 11 and 13 microM, respectively. When the assay method was applied to HTC cell samples, the SSAT activity, even at the control level, could easily be detected in as few as 20 microg protein of cell extract corresponding to 1 x 10(5) cells per determination, and 100 samples could be analyzed overnight. Thus, our HPLC method is a rapid and sensitive assay for the measurement of SSAT activity.


Effect of dopamine agents on schedule- and deprivation-induced drinking in rats.
The dopamine agonist, apomorphine, or its antagonist, haloperidol, was administered to rats whose drinking was induced by fixed-interval schedules of pellet delivery or by water deprivation. The first study revealed that both drugs produced dose-dependent decreases in bar-pressing and schedule-induced polydipsia (SIP). At higher doses, haloperidol also depressed the rate of pellet delivery. The second study demonstrated that the suppression in SIP obtained in the first study was primarily due to the direct effect of the drugs and not to changes they produced on the underlying food reinforcement schedule. The third study showed that both drugs suppressed water deprivation-induced drinking during a ten-minute session. Apomorphine delayed the onset of drinking, while haloperidol accelerated the cessation of drinking. The results indicated that apomorphine produced motor deficits that interfered with consummatory behavior, and that haloperidol interfered with the sensory feedback necessary to sustain consummatory behavior.


Structural and functional characterization of the GalNAc/Gal-specific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary.
The lectin found in mycelium and sclerotes of the phytopathogenic fungus Sclerotinia sclerotiorum is a homodimer consisting of two identical non-covalently bound subunits of 16,000 Da. CD spectra analysis revealed that the S. sclerotiorum agglutinin (SSA) contains predominantly beta-sheet structures. SSA exhibits specificity towards GalNAc whereby the hydroxyls at positions 4 and 6 of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of SSA can also accommodate disaccharides. The N-terminal sequence of SSA shares no significant similarity with any other protein except a lectin from the Sclerotiniaceae species Ciborinia camelliae. A comparison of SSA and the lectins from C. camelliae and some previously characterized lectins indicates that the Sclerotiniaceae lectins form a homogeneous family of fungal lectins. This newly identified lectin family, which is structurally unrelated to any other family of fungal lectins, is most probably confined to the Ascomycota.


Contribution of the IGCR1 regulatory element and the 3'Igh CTCF-binding elements to regulation of Igh V(D)J recombination.
Immunoglobulin heavy chain variable region exons are assembled in progenitor-B cells, from VH, D, and JH gene segments located in separate clusters across the Igh locus. RAG endonuclease initiates V(D)J recombination from a JH-based recombination center (RC). Cohesin-mediated extrusion of upstream chromatin past RC-bound RAG presents Ds for joining to JHs to form a DJH-RC. Igh has a provocative number and organization of CTCF-binding elements (CBEs) that can impede loop extrusion. Thus, Igh has two divergently oriented CBEs (CBE1 and CBE2) in the IGCR1 element between the VH and D/JH domains, over 100 CBEs across the VH domain convergent to CBE1, and 10 clustered 3'Igh-CBEs convergent to CBE2 and VH CBEs. IGCR1 CBEs segregate D/JH and VH domains by impeding loop extrusion-mediated RAG-scanning. Downregulation of WAPL, a cohesin unloader, in progenitor-B cells neutralizes CBEs, allowing DJH-RC-bound RAG to scan the VH domain and perform VH-to-DJH rearrangements. To elucidate potential roles of IGCR1-based CBEs and 3'Igh-CBEs in regulating RAG-scanning and elucidate the mechanism of the ordered transition from D-to-JH to VH-to-DJH recombination, we tested effects of inverting and/or deleting IGCR1 or 3'Igh-CBEs in mice and/or progenitor-B cell lines. These studies revealed that normal IGCR1 CBE orientation augments RAG-scanning impediment activity and suggest that 3'Igh-CBEs reinforce ability of the RC to function as a dynamic loop extrusion impediment to promote optimal RAG scanning activity. Finally, our findings indicate that ordered V(D)J recombination can be explained by a gradual WAPL downregulation mechanism in progenitor-B cells as opposed to a strict developmental switch.


[Age-related variation of adductor weight in the japanese scallop Mizuhopecten yessoensis (Jay) depending on cultivation technique].
Age-related variation in the absolute and relative adductor weight has been studied for 8 years in six samples of Japanese scallops Mizuhopecten yessoensis (Jay) of different generations from a shellfish farm in Alekseev Bight, Sea of Japan. The test character has proved to vary widely within the samples, with variation in shell size being relatively low. It has been shown that the dependence of adductor weight on the age of the scallop is not absolute: even-aged samples from different generations may differ significantly in this character. It is assumed that this effect is due to the influence of environmental factors on the development of scallops at early stages, which is especially strong under conditions of dense spat planting in the course of cultivation.


Frozen RPMI 8402 cells: a reliable source of terminal deoxynucleotidyl transferase positive control material.
A reliable source of positive control material is needed for immunoperoxidase or immunofluorescent terminal deoxynucleotidyl transferase (TdT) assays. The cell line RPMI 8402 was obtained from a patient with acute lymphoblastic leukemia and is positive for nuclear TdT. The kinetics of TdT expression in a series of cultured RPMI 8402 cells and the effect of cryopreservation on TdT expression were evaluated. The highest percentage of TdT+ cells was detected on days 5-8 of RPMI 8402 cell cultures. TdT positivity was preserved for more than 3 years in cells stored in suspension at -70 degrees C, and at least 76 days on slide preparations when fixed with methanol prior to freezing. We conclude that when RPMI 8402 cells with maximum TdT positivity are stored at -70 degrees C, aliquots can be used as a constant source of positive control material thereby eliminating the necessity and expense of a continuing culture system.


Utilization of the platelet release reaction in the blood compatibility assessment of polymers.
The release reaction is directly associated with platelet adhesion and aggregation, which are primary events leading to thrombus formation following contact of blood with artificial surfaces. This investigation examined the release reaction from the alpha granules of platelets after blood-polymer interaction, and utilized the measurement of beta-thromboglobulin (BTG), a platelet-specific protein, in the assessment of the in vitro blood compatibility of polymers. A radioimmunoassay was used, to determine the release of BTG following contact of blood with tubes of siliconized glass and polypropylene and flat sheets of poly(vinyl chloride) and silicone rubber. Polypropylene tubes caused less release of BTG than those of siliconized glass and silicone rubber induced less BTG release than poly(vinyl chloride). The investigation indicates a role for BTG measurement in blood compatibility assessment.


Supernumerary ring chromosomes in five bone and soft tissue tumors of low or borderline malignancy.
Five tumors (two myxoid malignant fibrous histiocytoma, two dermatofibrosarcoma protuberans, and one parosteal osteosarcoma) with ring chromosomes as the sole cytogenetic anomaly or as the only structural rearrangement were observed in a series of 60 karyotypically abnormal, nonlipogenic bone and soft tissue tumors (BST). All five tumors were of borderline or low malignancy. These findings support the suggestion that supernumerary ring chromosomes as the sole structural chromosomal aberration are not associated with any particular histopathologic diagnosis but may characterize a group of BST of borderline or low malignancy.


The relationship between financial strain, perceived stress, psychological symptoms, and academic and social integration in undergraduate students.
Financial strain may directly or indirectly (i.e., through perceived stress) impact students' psychological symptoms and academic and social integration, yet few studies have tested these relationships. The authors explored the mediating effect of perceived stress on the relationship between financial strain and 2 important outcomes: psychological symptomology and academic and social integration. Participants were 157 undergraduate students. Data were collected from December 2013 to March 2014. Cross-sectional data collection conducted using online survey software. It was found that perceived stress mediated the relationship between financial strain and (a) psychological symptomology and (b) academic and social integration. Both models included first-generation status as a covariate. Results suggest that perceived stress is an important intervention target for reducing psychological symptoms and improving academic and social integration for undergraduate students. Implications for university health centers and mental health professionals include incorporating a public health model to minimize stress risk.


'I think I will need help': A systematic review of who facilitates the recovery from gender-based violence and how they do so.
A key to advancing the eradication of gender-based violence (GBV) is knowing how to recover from it. To identify the changes that are indicators of having overcome GBV and determine the role of various support networks. We systematically searched Web of Science. Publications whose abstracts contained a minimum of two of the following words: (a) support or network or solidarity, (b) violence and (c) recovery or healing. Of the 273 documents retrieved, 52 were used using a narrative synthesis approach. For recovery, women must reconnect with themselves, with their environment and with the world in general. Doing so requires support from both formal and informal networks. The intervention of other people is a common element in successful recovery processes. We identify three requirements for the intervention of these support networks to be effective: not blaming the victim, making women part of their own recovery process by showing them their own transformation potential and promoting reflection on the socialization they have experienced in their affective-sexual relationships. There are multiple benefits to having formal or informal support during recovery. In informal networks, raising awareness and providing training are insufficient for promoting active support. Instead, the Second Order of Sexual Harassment must specifically be combatted. From the authors' previous research, harassed women and survivors have underlined the necessity to identify indicators of recovery and which kind of support has an impact on it.


[Contrastless venography in the diagnosis of varicosities of the subcutaneous veins of the lower extremities].
The results of the use of non-contrast venography in a surgical clinic on 38 patients with chronic venous defficiency and on 45 patients suspected of varicosis are generalized in this article. The obtained findings allow to recommend this method as a part of a complex examination of patients with various venous pathology of the lower extremities. Non-contrast venography method is described in this paper.


Genetic basis for multimodal relationship between apolipoprotein (a) size and lipoprotein (a) concentration in Mexican-Americans.
The reported general inverse relationship between apolipoprotein (a) (apo(a)) size and plasma lipoprotein (a) (Lp(a)) concentrations was further characterized using 927 samples taken from members of 42 Mexican-American families. When all samples were displayed in a scatter plot of apo(a) size versus natural log of Lp(a) concentration, the expected inverse relationship was observed (r2 = 0.24). However, the scatter plot revealed a multimodal pattern with at least two distinct modes of the inverse relationship between apo(a) size and Lp(a) concentration. Plots of 148 single-banded samples also showed the multimodal pattern, indicating that this pattern did not result from a difference between double- and single-banded samples. Also measured was the Lp(a) concentration associated with each of 508 apo(a) isoforms in samples from 254 double-banded phenotype individuals. These separated isoforms also showed the multimodal pattern. Using pedigree information, 29 different alleles were identified which were found in three or more family members. About 85% of variation in Lp(a) was explained by allele information, suggesting a genetic basis for the multimodal pattern. Thus, the data demonstrate at least two series of apo(a) alleles that have distinct relationships between apo(a) size and Lp(a) concentration.


Leonotis Leonurus improves the crosstalk between peripheral tissues both in vivo and in vitro.
Unravelling the anti-diabetic mechanism of action of L. leonurus at adipose, liver, muscle and pancreatic level. To investigate the mechanism of action of an organic extract of L. leonurus and marrubiin at the gene level in adipose, liver and muscle tissues of an obese rat model and in a co-culture model. Obese Wistar rats were fed a cafeteria diet for eight weeks, treated with an extract of L. Leonurus, marrubiin, sulfonylurea and aspirin for two weeks and the level of gene expression of selected markers were investigated across different tissues. The effects mediated by the different treatments were investigated in co-culture cell models involving 3T3-L1 (fat), Chang (liver), C2C12 (muscle) and INS-1 (pancreatic) cells under both normal and hyperglycemic conditions. L. leonurus extract mediated a significant increase in PPAR gamma, glucokinase, FAS and UCP2 gene expression in adipose tissue, whilst the opposite was observed in the liver. At the muscle level, a significant increase in FAS gene expression was observed relative to the obese control rats. Furthermore, the extract as well as marrubiin, modulated improvements in the adipokine profile. The co-culture models showed that the effect mediated by the extract was dependent on, the tissue type as well as the glycemic conditions. L. Leonurus extract as well as marrubiin exhibit anti-diabetic properties where the mechanism of action is mainly at the adipose tissue level. The increase in expression of the genes of interest mentioned above potentially play a protective role towards the liver and possibly towards the muscle tissues as well.


Voltage-gated calcium channels in developing GnRH-1 neuronal system in the mouse.
Migration of gonadotropin-releasing hormone-1 (GnRH-1) neurons from the nasal placode into the central nervous system occurs in all vertebrates. This study characterizes the expression of L- and N-type voltage-gated calcium channels (VGCCs) in migrating GnRH-1 neurons in mice. Class C (L-type) and class B (N-type) VGCGs were detected in GnRH-1 cells and cells in the olfactory and vomeronasal epithelium during prenatal development. This expression pattern was mimicked in a nasal explant model known to retain many characteristics of GnRH-1 development in vivo. Using this in vitro system, perturbation studies were performed to elucidate the role of VGCCs in GnRH-1 neuronal development. This report shows that olfactory axon outgrowth and GnRH-1 neuronal migration are attenuated when nasal explants are grown in calcium-free media, and that this effect is temporally restricted to an early developmental period. Blockade of either the L- or the N-type channel did not alter GnRH-1 cell number or overall olfactory axon outgrowth. However, blockade of N-type channels altered the distribution of GnRH-1 neurons in the periphery of the nasal explants. In these explants, more GnRH-1 neurons were located proximal to, and fewer GnRH-1 neurons distal to, the main tissue mass, suggesting a general decrease in the rate of GnRH-1 neuronal migration. These results indicate that extracellular calcium is required for initiating GnRH-1 neuronal migration and that these events are partially dependent on N-type VGCC signals.


Differential Implications of CSF3R Mutations in t(8;21) and CEBPA Double Mutated Acute Myeloid Leukemia.
Few data are available exploring mutations of the colony-stimulating factor 3 receptor (CSF3R) in acute myeloid leukemia (AML) in an all-round and systematic manner. The purpose of this study was to analyze the CSF3R mutations (CSF3Rmut) in AML with recurrent genetic abnormalities for potential synergistic pathomechanism. We retrospectively screened 1102 adult de novo AML patients with available next-generation sequencing (NGS) information on 132 genes related to hematologic disorders. The χ2, Mann-Whitney U tests were used to analyze their associations with clinicopathologic characteristics, and a propensity score matching (PSM) followed by Kaplan-Meier method was applied to measure their prognostic effects. Overall, CSF3Rmut were detected in 40 (3.6%) of 1102 patients with adult de novo AML. CSF3Rmut were predominantly enriched in AML with the CEBPA double mutations (CEBPAdm) (16/122, 13.1%), t(8;21) (12/186, 6.5%) and mutated RUNX1 (3/50, 6.0%), respectively. The CSF3Rmut loci and types differed according to AML subtypes, with frameshift-indels and premature stop confined in the t(8;21) AML [10/12 (83.3%)], and missense recurrently aggregated in the CEBPAdm AML [16/16 (100%)]. Cases with CSF3Rmut had a lower WBC count versus those with CSF3R wild-type (CSF3Rwt) in the t(8;21) AML cohort, with a borderline significance [median 5.45 (range 0.94-20.30) × 109/L) vs. 8.80 (range 0.96-155.00) × 109/L, P = .046]. CSF3Rmut were non-significantly associated with higher WBC counts [median 33.6 (range 6.8-287.6) × 109/L vs. 18.1 (range 1.7-196.0) × 109/L, P = .156] and significantly with lower immunophenotypic CD15 positivity [0/8 (0%) vs. 44/80 (55%), P = .009] as compared to CSF3Rwt in the CEBPAdm AML cohort. After propensity score matching followed by Kaplan-Meier analysis, CSF3Rmut cases had comparable disease-free survival (DFS) and overall survival (OS) to those with CSF3Rwt (P = .607 and P = .842, respectively) in the t(8;21) AML cohort. By contrast, CSF3Rmut showed an inclination towards inferior DFS compared to CSF3Rwt in the CEBPAdm AML cohort [median DFS 19.8 (95%CI 3.1-36.5) months vs. not reached (NR), P = .086]. No significant difference was found for OS between CSF3Rmut and CSF3Rwt cases (P = .943). We concluded that CSF3Rmut were frequently enriched in patients with t(8;21) and CEBPAdm subtypes among AML, but showed divergent clinicopathologic features, mutation loci and types and differing prognostic aspects.


Progression-free Survival After Second Line of Therapy for Metastatic Clear Cell Renal Cell Carcinoma in Patients Treated with First-line Immunotherapy Combinations.
Immunotherapy (IO)-based combinations used to treat metastatic clear cell renal cell carcinoma (ccRCC) include dual immune checkpoint inhibition with ipilimumab and nivolumab (IO/IO) and several combinations of vascular endothelial growth factor receptor-targeting tyrosine kinase inhibitors (TKIs) with an immune checkpoint inhibitor (TKI/IO). IO/IO and TKI/IO approaches have not been compared directly, and it is unknown whether patients who do not respond to first-line IO/IO can salvage long-term survival by receiving a second-line TKI. Progression-free survival after second-line therapy (PFS-2) evaluates the ability to be salvaged by second-line therapy. We retrospectively evaluated 173 patients treated with first-line IO/IO or TKI/IO for metastatic ccRCC at Memorial Sloan Kettering Cancer Center and report PFS-2, overall survival, and response to second line of therapy (ORR2nd) for groups defined by first-line category. Although ORR2nd was significantly higher with IO/IO than with TKI/IO (47% vs 13%, p < 0.001), there was no significant difference in median PFS-2 for TKI/IO versus IO/IO (44 vs 23 mo, log-rank p = 0.1) or restricted mean survival time (RMST) for PFS-2 when adjusted for propensity score (33 vs 30 mo; difference 2.6 mo [95% confidence interval {CI}: -2.6, 7.9]; p = 0.3). There was also no significant difference in RMST for overall survival when adjusted for propensity score (38 vs 37 mo; group difference 1.0 mo [95% CI: -3.4, 5.5]; p = 0.7). These findings do not support a change in current utilization practices for IO/IO and TKI/IO treatment strategies for ccRCC. PATIENT SUMMARY: In cases of metastatic clear cell renal cell carcinoma, no significant difference was observed in progression-free survival after second line of therapy between patients receiving ipilimumab plus nivolumab and those receiving a combination of a tyrosine kinase inhibitor and an immune checkpoint inhibitor.


Is there an EKC between economic growth and air pollutant emissions in SAARC countries? Evidence from disaggregated analysis.
The manufacturing and construction (M&C) sector not only plays a vital role in promoting economic growth, but is also a significant contributor to global air pollution. Growing concerns regarding air pollutant emissions necessitate a more disaggregated (i.e., sectoral) investigation in order to identify the major contributors. This study employs aggregated and disaggregated data to determine the fundamental effects of economic growth (i.e., overall growth and sectoral growth) on air pollutant emissions (APE) (specifically, PM2.5 and PM10 released by the M&C sector) in SAARC economies between 1995 and 2018. It assesses the environmental Kuznets curve (i.e., inverted U-shaped and N-shaped) using the feasible generalized least squares (FGLS), panel-corrected standard errors (PCSE), and generalized method of moments (GMM) techniques. The sectoral analysis reveals the presence of an N-shaped EKC while the overall analysis indicates an inverted U-shaped EKC. Population, financial development (FD), and merchandise exports (MX) have no influence on the estimates. Population and FD increase APE in all models, whereas the effects of MX vary between models. As SAARC economies are capital-deficient, these economies can adopt unbalanced environmental protection policies. First, focus on major contributing sectors (e.g., M&C sector) to curb APE, then focus on less emitting sectors in turn. By implementing pollution reduction strategies on M&C sector activities, governments may reach their threshold (peak) points earlier than expected. A reduction in APE is impossible without rigorous monitoring and application. Being capital-deficient nations and given the collective nature of the problem, a Transboundary Haze/Pollution agreement is required to solve this issue.


A novel route to estimate the photocatalytic activity of titanium dioxide via voltage measurements.
Titanium dioxide is known as a photocatalyst, that may be activated by UV-A light and thus be able to lead to chemical reactions, to mineralize organic compounds, to inactivate biomolecules and to kill microorganisms, respectively. To estimate the capability of the photocatalytic activity a novel experimental setup using photovoltage measurements was studied. A distinct correlation between the photocatalytic activity of the titanium dioxide samples and the measurable photovoltages could be demonstrated. The experimental setups included the construction of different electrochemical cells based on TiO2 and using different liquid and gel electrolytes. The investigations were carried out on titanium dioxide layers as well as on TiO2 particle materials. The electrochemical measurement of the resulting voltage was optimised and the results were compared with conventionally used methods for the determination of the photocatalytic activity like the decolourization of methylene blue solutions. Additionally, the increase of carbon dioxide concentration in the gas phase was taken as a measure for total mineralization. Results indicate strong correlations between the different methods.


First Evidence of Atrial Conduction Disorders in Pediatric Patients With Congenital Heart Disease.
This study sought to investigate whether pediatric patients with congenital heart disease (CHD) already have atrial conduction disorders early in life. The authors conducted first-in-children epicardial mapping in 10 pediatric patients with CHD undergoing primary open heart surgery. Areas of conduction delay (CD) and block (CB) were present in all patients and were particularly observed at Bachmann's bundle (CD: 4.9%; CB: 2.3%), followed by the right atrium (CD: 3.7%; CB: 1.6%) and, to a lesser degree, the left atrium (CD: 1.8%; CB: 1.0%). Conduction abnormalities may by aggravated over time (e.g., aging, residual lesions, or valvular dysfunction), predisposing these patients to atrial arrhythmias early in life.


An outcome evaluation of the SOS Suicide Prevention Program.
We examined the effectiveness of the Signs of Suicide (SOS) prevention program in reducing suicidal behavior. Twenty-one hundred students in 5 high schools in Columbus, Ga, and Hartford, Conn, were randomly assigned to intervention and control groups. Self-administered questionnaires were completed by students in both groups approximately 3 months after program implementation. Significantly lower rates of suicide attempts and greater knowledge and more adaptive attitudes about depression and suicide were observed among students in the intervention group. The modest changes in knowledge and attitudes partially explained the beneficial effects of the program. SOS is the first school-based suicide prevention program to demonstrate significant reductions in self-reported suicide attempts.


A complex 'mRNA degradation code' controls gene expression during animal development.
Current understanding of the molecular mechanisms underlying mRNA degradation indicates that specific mRNA degradation rates are primarily encoded within the mRNA message itself in the form of cis-regulatory elements bearing particular primary sequences and/or secondary-structures. Such control elements are operated by RNA-binding proteins (RBPs) and/or miRNA-containing complexes. Based on the large number of RBPs and miRNAs encoded in metazoan genomes, their complex developmental expression and that specific RBP and miRNA interactions with mRNAs can lead to distinct degradation rates, I propose that developmental gene expression is shaped by a complex 'mRNA degradation code' with high information capacity. Localised cellular events involving the modification of RBP and/or miRNA target sequences in mRNAs by alternative polyadenylation added to the activation of specific RBP and miRNA activities via cell signalling are predicted to further expand the capacity of the mRNA degradation code by coupling it to dynamic events experienced by cells at specific spatiotemporal coordinates within the developing embryo.


Feasibility and preliminary efficacy of a novel web-based physical activity intervention in adults with overweight/obesity: A pilot randomized controlled trial.
Only 14% of adults with obesity attain federal guidelines for physical activity (PA), but few interventions address obesity-specific barriers to PA. We designed the web-based Physical Activity for The Heart (PATH) intervention to address this gap. Test the feasibility and preliminary efficacy of PATH for promoting PA and reducing cardiovascular disease (CVD) risk in adults with overweight/obesity. In a 12-week pilot RCT, participants were randomized to PATH (n = 41) or wait-list control (n = 41) groups. Treatment group received access to PATH and met twice/month with a remote coach. The control group received a self-help PA guide and newsletters on general health. Moderate-to-vigorous PA (MVPA) was assessed via Actigraph-GT3X, steps via Fitbit Charge 2™, weight via smart scale, blood pressure (BP) via Omron BP device, and lipids/HbAIC via dry blood spot. Linear mixed modeling examined between- and within-group differences in PA and CVD risk. The sample (N = 82) was on average 55.9 ± 8.2 years old; mean BMI 35.5 ± 6.2 kg/m2; 57.3% white and 80.5% female. Recruitment lasted 6-months, and 12-week retention was 96.3%. Treatment group accessed PATH ≥twice/week (92.1%), spent ≥10 min/visit (89.5%) and thought the site was culturally appropriate (79%). At 12 wks, the PATH group had greater mean changes in weekly MVPA (+58.9 vs. +0.9 min, p = .024) and daily steps (+1246.4 vs. -64.2 steps, p = .002) compared to the control group. Also, the PATH group improved in weight, BMI, body fat, waist circumference, and BP (p < .05). The PATH intervention is feasible/acceptable and demonstrated preliminary efficacy for promoting PA among adults with overweight/obesity.


Hearing Status and Static Postural Control of Collegiate Athletes.
Because of the close proximity of the cochlea, vestibular apparatus, and shared neurovascular structures, the static postural control of athletes who are deaf or hard of hearing (D/HoH) may be different from that of athletes who are hearing. Limited research is available to quantify differences between these athletes. To determine the effect of hearing status and stance condition on the static postural control of athletes. Cross-sectional study. Athletic training facilities. Fifty-five collegiate varsity athletes who were D/HoH (age = 20.62 ± 1.80 years, height = 1.73 ± 0.08 m, mass = 80.34 ± 18.92 kg) and 100 university club athletes who were hearing (age = 20.11 ± 1.59 years, height = 1.76 ± 0.09 m, mass = 77.66 ± 14.37 kg). Participants completed the Modified Clinical Test of Sensory Interaction and Balance on a triaxial force plate. Anteroposterior and mediolateral (ML) center-of-pressure (CoP) velocity, anteroposterior and ML CoP amplitude root mean square, and 95% ellipse sway area were calculated. Athletes who were D/HoH had a larger CoP velocity, larger ML root mean square, and larger sway area than those who were hearing (P values < .01). A significant main effect of stance condition was observed for all postural control variables (P values < .01). During the Modified Clinical Test of Sensory Interaction and Balance, athletes who were D/HoH demonstrated a larger sway area compared with athletes who were hearing. Therefore, individualized baseline assessments of static postural control may be warranted for athletes who are D/HoH as opposed to comparisons with existing normative data.


Implications of Bit1 and AIF overexpressions in esophageal squamous cell carcinoma.
Overwhelming evidence has demonstrated that Bit1 and AIF as mitochondrial proteins are implicated in the development and progression of a variety of tumors. However, their expressions and biological functions in esophageal squamous cell carcinoma (ESCC) remain to be delineated. In the present study, we found that Bit1, AIF, and Bcl-2 levels in ESCC tissues were significantly higher than those in normal esophageal epithelial tissues and dysplasia tissues (P < 0.05). Stepwise investigation demonstrated that Bit1 and Bcl-2 levels were both tightly associated with lymphatic metastasis and TNM staging (P < 0.05), and the levels of Bit1 mRNA as well as AIF and Bcl-2 proteins were both closely related to tumor differentiation (P < 0.05), but not related to the patients' age and gender (P > 0.05). Importantly, Bit1 mRNA and protein levels in ESCC with lymphatic metastasis and TNM staging in III and IV were markedly higher than that without lymphatic metastasis and TMN staging in I and II. Further analysis showed that expression of Bit1 protein was both positively correlated with expressions of AIF and Bcl-2 proteins (r = 0.408 and 0.405, respectively; P < 0.05). Correctively, our data cited earlier suggest that Bit1 plays pivotal roles in the development and progression of ESCC, and its biological functions in ESCC may be closely associated with AIF and Bcl-2 levels.


Discordant GH and IGF-1 Results in Treated Acromegaly: Impact of GH Cutoffs and Mean Values Assessment.
Discordant growth hormone (GH) and insulin-like growth factor-1 (IGF-1) values are frequent in acromegaly. To evaluate the impact of different GH cutoffs on discordance rate. To investigate whether the mean of consecutive GH measurements impacts discordance rate when matched to the last available IGF-1 value. Retrospective study. Referral center for pituitary diseases. Ninety acromegaly patients with at least 3 consecutive evaluations for GH and IGF-1 using the same assay in the same laboratory (median follow-up 13 years). Multimodal treatment of acromegaly. Single fasting GH (GHf) and IGF-1 (IGF-1f). Mean of 3 GH measurements (GHm), collected during consecutive routine patients' evaluations. At last evaluation GHf values were 1.99 ± 2.79 µg/L and age-adjusted IGF-1f was 0.86 ± 0.44 × upper limit of normality (mean ± SD). The discordance rate using GHf was 52.2% (cutoff 1 µg/L) and 35.6% (cutoff 2.5 µg/L) (P = 0.025). "High GH" discordance was more common for GHf <1.0 µg/L, while "high IGF-1" was predominant for GHf <2.5 µg/L (P < 0.0001). Using GHm mitigated the impact of GH cutoffs on discordance (GHm <1.0 µg/L: 43.3%; GHm <2.5 µg/L: 38.9%; P = 0.265). At receiver-operator characteristic curve (ROC) analysis, both GHf and GHm were poor predictors of IGF-1f normalization (area under the curve [AUC] = 0.611 and AUC = 0.645, respectively). The prevalence of disease-related comorbidities did not significantly differ between controlled, discordant, and active disease patients. GH/IGF-1 discordance strongly depends on GH cutoffs. The use of GHm lessen the impact of GH cutoffs. Measurement of fasting GH levels (both GHf and GHm) is a poor predictor of IGF-1f normalization in our cohort.


Validation of the Kihon Checklist and the frailty screening index for frailty defined by the phenotype model in older Japanese adults.
The term "frailty" might appear simple, but the methods used to assess it differ among studies. Consequently, there is inconsistency in the classification of frailty and predictive capacity depending on the frailty assessment method utilised. We aimed to examine the diagnostic accuracy of several screening tools for frailty defined by the phenotype model in older Japanese adults. This cross-sectional study included 1,306 older Japanese adults aged ≥ 65 years who underwent physical check-up by cluster random sampling as part of the Kyoto-Kameoka Study in Japan. We evaluated the diagnostic accuracy of several screening instruments for frailty using the revised Japanese version of the Cardiovascular Health Study criteria as the reference standard. These criteria are based on the Fried phenotype model and include five elements: unintentional weight loss, weakness (grip strength), exhaustion, slowness (normal gait speed), and low physical activity. The Kihon Checklist (KCL), frailty screening index (FSI), and self-reported health were evaluated using mailed surveys. We calculated the non-parametric area under the receiver operating characteristic curve (AUC ROC) for several screening tools against the reference standard. The participants' mean (standard deviation) age was 72.8 (5.5) years. The prevalence of frailty based on the Fried phenotype model was 12.2% in women and 10.3% in men. The AUC ROC was 0.861 (95% confidence interval: 0.832-0.889) for KCL, 0.860 (0.831-0.889) for FSI, and 0.668 (0.629-0.707) for self-reported health. The cut-off for identifying frail individuals was ≥ 7 points in the KCL and ≥ 2 points in the FSI. Our results indicated that the two instruments (KCL and FSI) had sufficient diagnostic accuracy for frailty based on the phenotype model for older Japanese adults. This may be useful for the early detection of frailty in high-risk older adults.


The effects of viscoelastic parameters on residual stress development in a zirconia/glass bilayer dental ceramic.
The aim of this study was to test the hypothesis that the residual stresses in a zirconia-based bilayer dental composite system can be tailored through heat treatment above and below the glass transition temperature of glass veneers. Ceramic bilayer disc specimens were prepared from a zirconia core and a glass veneer. Each bilayer ceramic group was heat treated 40 degrees C below, 20 degrees C and 40 degrees C above and at the glass transition temperature of the glass veneer, and cooled using a fast or a slow cooling rate. Specimens were tested for flexure strength using a biaxial bending fixture. Residual stresses were calculated using a fracture mechanics approach. Heat treatments produced significant differences (p < or =0.05) between the mean flexural strengths of the heat treatment groups when the specimens were cooled using a fast cooling rate. However, there was not a significant difference (p >0.05) between the mean flexural strengths of the heat treatment groups when a slow cooling rate was used. Fractures initiated from the veneer surfaces of the specimens. Heat treatment above and below the glass transition temperature of the veneer layer, and the cooling rate have a significant effect on the flexural strength of the bilayer ceramic laminates. The existence of residual compressive stress is the most likely reason for the observed strength increases. Residual stresses can be modified using the elastic-viscoelastic relaxation behavior of a glass veneer.


Comparison of mid-term clinical outcomes after acute myocardial infarction in diabetic men between living alone and living together.
Because living alone is associated with an increased risk of type 2 diabetes mellitus in men but not women, living alone may be a risk factor of cardiovascular events after acute myocardial infarction (AMI) in diabetic men. The aim of the present study was to investigate the association between living alone and mid-term clinical outcomes after AMI in diabetic men. We conducted a single center, retrospecitve study. The primary endpoint was the major adverse cardiovascular events (MACE) defined as the composite of all cause death, AMI, and target vessel revascularization. A total of 253 AMI men with diabetes mellitus were included from our hospital records, and divided into the living together group (n = 203) and the living alone group (n = 50). Median follow-up duration was 239 days (Q1: 94 days, Q3: 451 days). A total of 66 MACE was observed during the study period, and Kaplan-Meier curves were constructed to compare the MACE. The MACE was more frequently observed in the living alone group than the living together group (P = 0.041). Multivariate Cox regression analysis revealed that the living alone group was significantly associated with the MACE (Odds ratio: 1.770, 95% confidence interval 1.018-3.077, P = 0.043) after known clinical risk factors. In conclusion, living alone was significantly associated with the mid-term MACE after AMI in diabetic men. It may be important to provide multiple interventions including lifestyle guidance as well as sufficient acute medical care for such high-risk patients.


A phase I clinical trial of weekly oral topotecan for relapsed small cell lung cancer.
To determine the dose-limiting toxicities (DLTs) and the maximum tolerated dose (MTD) of oral topotecan administered weekly in patients with relapsed small cell lung cancer (SCLC). Patients were treated with oral topotecan on days 1, 8, and 15, every 28 days. The dose was escalated by 0.5 mg/m² increments from the starting dose of 3 mg/m² until the MTD was reached. DLTs were defined as grade 4 neutropenia, febrile neutropenia, grade 4 thrombocytopenia, non-hematologic toxicity ≥grade 3, any toxicity precluding the treatment on days 8 or 15 of the first cycle, or delay of the second cycle for more than 7 days. Eighteen patients were enrolled. Thirteen patients received oral topotecan as second-line and five as third- or further-line treatment. The DLT level was reached at 4.5 mg/m², and the MTD was determined to be 4 mg/m². DLTs consisted of grade 2/3 neutropenia and grade 2 thrombocytopenia precluding treatment on day 15 of the first cycle or on day 1 of the second cycle. The most frequent toxicities were grade 2-3 neutropenia (27.8 % of patients), grade 2-3 anemia (33.3 %), grade 2 thrombocytopenia (16.7 %), and grade 2-3 fatigue (44.4 %). The response rate was 11.1 %, the median progression-free survival 2.3 months, and the median overall survival 5.1 months. The recommended phase II dose of weekly oral topotecan in pretreated patients with SCLC is 4 mg/m² on days 1, 8, and 15 every 28 days.


Hypertrophic cardiomyopathy. Clinical spectrum and treatment.
HCM is a heterogeneous disease genotypically, phenotypically, pathophysiologically, clinically, and therapeutically. In decisions on the management of these patients, it is important to recognize this heterogeneity and to direct therapy at the predominant abnormalities.


Bodily expression of psychological distress in adolescents: a qualitative study.
The bodily expression of psychological disorders is one of the leading motives for consultations in adolescent medicine. The diagnostic entity corresponding to DSM-5 "Somatic symptom and related disorders" is sparsely used in the English-language literature, especially for adolescents. Qualitative studies on this topic mostly concern the experiences of healthcare professionals. This study seeks to explore the experience of adolescents expressing psychological distress through their body. This exploratory research took place in a Paris hospital department of adolescent medicine. Our sampling method was purposive. For inclusion, patients had to be aged 11-24 years, with a "somatic symptom disorder" meeting the DSM-5 criteria. Semi-directive interviews were proposed with visual narration inspired by photoelicitation. Thematic analysis allowed us to explore the data with an inductive approach. Thirty adolescents were interviewed; they were 14-22 years old and mostly had somatic symptom or functional neurological disorders. Three principal themes emerged from our analysis of the interview contents: the personal, including bodily, experience of the disorder, the experience of relationships, and the question of what is visible through the body. This research allowed us to discuss the reversal of generations, the function of the DSM-5 diagnosis, illuminated by sociology, and finally, cultural pathways. It shows the importance of recognizing the reality of the adolescents' bodily feelings, reassuring them by ruling out serious causes, and supporting their search for meaning. It is important to think about a specific framework of family therapy that can make effective use of this experience of the reversal of generations.


Reflections from undergraduate teaching experiences: some problems and solutions of restoring teeth with dental resin composite instead of dental amalgam.
This is a simple personal reflection on some of the problems and solutions related to the increasing use of composite resin to replace dental amalgam when dental students learn to place restorations at the start of their careers. To the author, much seems common sense. Much is not, or cannot, be backed up by ideal science and some may still be considered outdated to the more progressive practitioner. Unfortunately, prospective, ideally-designed clinical trials may no longer be possible to seek the answers we lack due to ethical, organisational, financial or other constraints.


Structural mechanisms of TRPV6 inhibition by ruthenium red and econazole.
TRPV6 is a calcium-selective ion channel implicated in epithelial Ca2+ uptake. TRPV6 inhibitors are needed for the treatment of a broad range of diseases associated with disturbed calcium homeostasis, including cancers. Here we combine cryo-EM, calcium imaging, and mutagenesis to explore molecular bases of human TRPV6 inhibition by the antifungal drug econazole and the universal ion channel blocker ruthenium red (RR). Econazole binds to an allosteric site at the channel's periphery, where it replaces a lipid. In contrast, RR inhibits TRPV6 by binding in the middle of the ion channel's selectivity filter and plugging its pore like a bottle cork. Despite different binding site locations, both inhibitors induce similar conformational changes in the channel resulting in closure of the gate formed by S6 helices bundle crossing. The uncovered molecular mechanisms of TRPV6 inhibition can guide the design of a new generation of clinically useful inhibitors.


Use of flumazenil in intoxicated patients with coma. A double-blind placebo-controlled study in ICU.
In a double-blind placebo-controlled prospective clinical trial we studied the efficacy and safety of the benzodiazepine antagonist, flumazenil. In 23 patients admitted to the Intensive Care Unit with coma due to overdose with benzodiazepines or other sedatives, flumazenil i.v. (up to 2 mg or placebo) was given. In 13 patients given flumazenil the Glasgow Coma Scale (GCS) increased significantly from 4.9 to 7.8 (p less than 0.05). Six of these 13 patients, including mainly benzodiazepine mono-intoxications, needed only one series of injections (up to 1.0 mg flumazenil); the GCS increased thereby from 4.5 to 10.7 within a maximum of 5 min (p less than 0.01). In the remaining 7 patients, needing two series of injections of flumazenil (up to 2.0 mg), GCS did not rise significantly and coma was related to intoxications with nonbenzodiazepine sedatives, flunitrazepam and in one patient, encephalitis. In the 10 patients receiving placebo, the GCS did not change. A significant increase in the GCS from 5.5 to 10.8 (p less than 0.001) was, however, observed when flumazenil (up to 1.0 mg) was given after placebo. In patients with EEG monitoring the changes in waveform pattern paralleled the clinical response. Effects could be detected within 1-2 min after flumazenil injection and lasted up to 45 min. There were no adverse reactions or benzodiazepine withdrawal symptoms. We conclude that flumazenil is an effective and safe drug in the treatment of benzodiazepine overdose. The use of flumazenil is of diagnostic value in mixed-drug intoxications or coma of unknown origin and is of therapeutic importance for reversal of benzodiazepine intoxications.


In situ small-angle neutron scattering study of nitrogen adsorption and condensation in mesoporous silica glass CPG-10-75.
Film formation and capillary condensation of nitrogen at 78 K on the mesoporous controlled pore glass CPG-10-75 have been studied at certain relative pressures by in situ small-angle neutron scattering. On desorption ramified clusters of vapor filled voids have been observed, but not on adsorption. The kinetics of adsorption and desorption have been followed. The experimental results are discussed with respect to recent theoretical studies of fluids in complex pore systems.


Anorexia nervosa: selective processing of food-related word and pictorial stimuli in recognition and free recall tests.
Maladaptive processing of food cues is considered pivotal in the psychopathology of anorexia nervosa. However, the influence of hunger and differences in processing because of the type of stimuli remain largely unclear. Memory bias for food-related pictorial and semantic stimuli was assessed in a recognition and a free recall test in 16 anorexia nervosa (AN) patients, 16 control participants with food intake prior to the study (CG-FI) and 16 control participants with a fasting period prior to the study (CG-NF). Compared with CG-FI participants, both AN and CG-NF participants responded faster to food-related as compared with neutral words (p < .001) in the recognition test. Differences were found for word but not for pictorial stimuli. No group differences were observed with respect to the number of correct retrievals in either the recognition or the free recall test. The present study found behavioral indications of abnormal processing of food-related and neutral stimuli in anorectic patients similar to those found in fasted healthy controls. Results are discussed in terms of self-schemata in eating disorders, competitive interference, and levels of processing.


Intake of red meat and heterocyclic amines, metabolic pathway genes and bladder cancer risk.
We analyzed the association between meat intake, heterocyclic amines (HCAs) and bladder cancer (BC) risk in a large case-control study comprised of 884 BC cases and 878 healthy controls, recruited from 1999 to 2009. Epidemiologic and dietary data were collected via an in-person interview. Compared to the lowest quartile of red meat intake, the odds ratios (ORs) for the second, third and fourth quartiles were 1.17 (95% CI: 0.87-1.58), 1.47 (95% CI: 1.09-1.99) and 1.95 (95% CI: 1.41-2.68), respectively, (p-for trend <0.001). In a subset of participants with intakes of HCAs available, compared with those with the lowest quartile of intake, the ORs for the second, third and fourth quartiles were 1.47 (95% CI: 0.60-3.64), 2.58 (95% CI: 1.09-6.11) and 3.32 (95% CI: 1.37-8.01), respectively, (p for trend <0.001). In cumulative analysis of SNPs in the pathway, compared with subjects carrying 0-4 unfavorable genotypes, subjects carrying 5 and 6 or more unfavorable genotypes were at 1.60-fold (95% CI: 1.20-2.12) and 2.37-fold (95% CI: 1.82-3.10) increased risk, respectively. Moreover, subjects carrying six or more unfavorable genotypes and whose red meat intake was in the highest quartile were at 5.09-fold increased risk (95% CI: 2.89-8.96; p < 0.001). These results strongly support that high red meat intake, high intake of HCAs and carrying high number of unfavorable genotypes in the HCA metabolic pathways are associated with increased risk of BC in the study population.


Ce-based catalysts used in advanced oxidation processes for organic wastewater treatment: A review.
Refractory organic pollutants in water threaten human health and environmental safety, and advanced oxidation processes (AOPs) are effective for the degradation of these pollutants. Catalysts play vital role in AOPs, and Ce-based catalysts have exhibited excellent performance. Recently, the development and application of Ce-based catalysts in various AOPs have been reported. Our study conducts the first review in this rapid growing field. This paper clarifies the variety and properties of Ce-based catalysts. Their applications in different AOP systems (catalytic ozonation, photodegradation, Fenton-like reactions, sulfate radical-based AOPs, and catalytic sonochemistry) are discussed. Different Ce-based catalysts suit different reaction systems and produce different active radicals. Finally, future research directions of Ce-based catalysts in AOP systems are suggested.


Biomarkers and molecular imaging in gastrointestinal cancers.
The best means to improve gastrointestinal cancer survival is screening and treatment of early lesions. In esophageal adenocarcinoma, it is believed that low-grade dysplasia and perhaps even high-risk Barrett's esophagus represent the most attractive targets for achieving a cure. An issue with Barrett's esophagus is that endoscopy alone cannot distinguish Barrett's esophagus from columnar-lined epithelium or from areas of low-grade dysplasia. Much effort, therefore, has been devoted to discover molecular biomarkers of high-risk states and to develop imaging tools for detecting these biomarkers in a manner that could assist real-time in vivo targeting of sites for biopsy. The strategy we have used is to generate stem cell clones from Barrett's esophagus biopsy specimens and to compare their gene expression profiles with patient-matched stem cell clones of the esophageal squamous epithelia and gastric cardia. It is anticipated that by mining the expression data sets of these Barrett's stem cell clones, we will be able to identify unique cell surface markers of the Barrett's stem cells against which cytotoxic antibodies or aptamers can be developed and used to aid the endoscopist in identifying regions of atypia for biopsy, perform a real-time diagnosis, stratify patients during the examination, and, ultimately, direct therapy in a preemptive manner.


Pelvic solitary fibrous tumor originally diagnosed as prostatic in origin.
A 71-year-old man was referred to our hospital because of intermittent urine stream and postmicturition dribbling. Magnetic resonance imaging (MRI) results suggested the mass to be a malignant mesenchymal tumor arising from the left lobe of the prostate, on the basis of the presence of a beak sign. Radical prostatectomy and partial rectal excision with subsequent colostomy were performed. Contrary to preoperative MRI, no prostate involvement was found on histologic examination. Histopathologic and immunohistochemical findings showed typical characteristics of solitary fibrous tumors. The patient's postoperative course was uneventful. He showed no signs of recurrence and metastasis at 2-year follow-up.


Development of a Multiplex Polymerase Chain Reaction (PCR) Assay for the Potential Detection of Insect Contaminants in Food.
Molecular methods can potentially be used to detect insect contaminants of food products. In this study, we used three sets of group-specific primers, two of them targeting the amplification of two regions of the insect's mitochondrial cytochrome c oxidase subunit I (COI-Fa and COI-Fb) and the other targeting a region of the nuclear protein-coding wingless (wg) gene. Using singleplex and multiplex polymerase chain reaction (PCR), we evaluated the three sets of primers using genomic DNA (gDNA) from 48 insect species including food storage insect pests and known vectors of foodborne pathogens. Seven plant-based food matrices were also evaluated for exclusivity testing. Additionally, we spiked fragments from five insect species in a selected food matrix (whole wheat flour). Singleplex and multiplex PCR amplified single specific bands (401-449 bp), corresponding to the wg gene, from insect species belonging to families Blattidae and Formicidae, and in Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae). The COI-Fa primers amplified specific bands (171-188 bp) in all Dipteran species and the COI-Fb primers amplified a specific band (∼140 bp) in DNA from Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and P. interpunctella. However, the presence of specific bands in most Coleopterans was not consistent. No amplicon bands were observed in any of the food matrixes tested and the expected pattern of amplicon bands was seen in multiplex reactions using gDNA from spiked food samples. Our multiplex PCR assay targeted specific groups of insects that commonly contaminate foods without amplifying bands from the food matrixes tested; thus, molecular methods may be suitable for detecting insects or their fragments in foods.


Clonal heterogeneity of cyclic AMP responsiveness: a comparison of malignant murine lymphoid cell lines.
Various murine lymphoid tumor cell lines were examined for their alterations in intracellular levels of cyclic AMP following exposure to several pharmacologic agents. The agents tested in the present study included 3-isobutyl-1-methylxanthine (MIX), isoproterenol (ISO), prostaglandin E2 (PGE), histamine, and cholera toxin (CT). B cell tumors were generally found to be less responsive to beta-adrenergic and PGE-induced increases in cyclic AMP than were T cell tumors. An exception to this general finding was the murine B lymphoma cell line WEHI-231, which demonstrated marked sensitivity to ISO and PGE. Macrophage tumors were generally found to be even less responsive to PGE than were the B cell tumors, again with some notable exceptions (P388.D1). Cholera toxin produced differential effects in B, T, and macrophage tumors, both in terms of the absolute magnitude of the cyclic AMP response and the kinetics of this response. A T lymphoma cell line (EL-4) was identified that seems to be totally unresponsive to cell surface receptor-mediated increases in intracellular levels of cyclic AMP. The results of this study are discussed in terms of the usefulness of murine lymphoid tumors for the study of pharmacologic modulation of the immune response by cyclic AMP-elevating agents. These results demonstrate the high degree of cellular heterogeneity that can exist within normal lymphoid populations of cells, and suggest that cloned cell lines may be useful in the biochemical characterization of subsets of lymphoid cells.


Assessing visual fields for driving in patients with paracentral scotomata.
The binocular Esterman visual field test (EVFT) is the current visual field test for driving in the UK. Merging of monocular field tests (Integrated Visual Field, IVF) has been proposed as an alternative for glaucoma patients. To examine the level of agreement between the EVFT and IVF for patients with binocular paracentral scotomata, caused by either ophthalmological or neurological conditions, and to compare outcomes with useful field of view (UFOV) performance, a test of visual attention thought to be important in driving. 60 patients with binocular paracentral scotomata but normal visual acuity (VA) were recruited prospectively. Subjects completed and were classified as "pass" or "fail" for the EVFT, IVF and UFOV. Good agreement occurred between the EVFT and IVF in classifying subjects as "pass" or "fail" (kappa = 0.84). Classifications disagreed for four subjects with paracentral scotomata of neurological origin (three "passed" IVF yet "failed" EVFT). Mean UFOV scores did not differ between those who "passed" and those who "failed" both visual field tests (p = 0.11). Agreement between the visual field tests and UFOV was limited (EVFT kappa = 0.22, IVF kappa 0.32). Although the IVF and EVFT agree well in classifying visual fields with regard to legal fitness to drive in the UK, the IVF "passes" some individuals currently classed as unfit to drive due to paracentral scotomata of non-glaucomatous origin. The suitability of the UFOV for assessing crash risk in those with visual field loss is questionable.


A Neural Mechanism for Sensing and Reproducing a Time Interval.
Timing plays a crucial role in sensorimotor function. However, the neural mechanisms that enable the brain to flexibly measure and reproduce time intervals are not known. We recorded neural activity in parietal cortex of monkeys in a time reproduction task. Monkeys were trained to measure and immediately afterward reproduce different sample intervals. While measuring an interval, neural responses had a nonlinear profile that increased with the duration of the sample interval. Activity was reset during the transition from measurement to production and was followed by a ramping activity whose slope encoded the previously measured sample interval. We found that firing rates at the end of the measurement epoch were correlated with both the slope of the ramp and the monkey's corresponding production interval on a trial-by-trial basis. Analysis of response dynamics further linked the rate of change of firing rates in the measurement epoch to the slope of the ramp in the production epoch. These observations suggest that, during time reproduction, an interval is measured prospectively in relation to the desired motor plan to reproduce that interval.


Angiotensin II reduces transport-dependent oxygen consumption but increases transport-independent oxygen consumption in immortalized mouse proximal tubular cells.
Oxidative stress is closely associated with renal dysfunction following diabetes and hypertension. Angiotensin II (Ang II) can activate the NADPH-oxidase, increasing oxidative stress that is thought to blunt proximal tubular electrolyte transport and thereby oxygen consumption (QO₂). We investigated the effect of Ang II on QO₂ in immortalized mouse proximal tubular cells over-expressing the NADPH oxidase subunit p22(phox); a model of increased oxidative stress. Cultured cells were exposed to either Ang II or H₂O₂ for 48 h. QO₂ was determined during baseline (113 mmol/l NaCl; transport-dependent QO₂) and during sodium-free conditions (transport-independent QO₂). Ang II reduced transport-dependent QO₂ in wild-types, but not in p22(phox) which also displayed increased QO₂ at baseline. Transport-independent QO₂ was increased in p22(phox) and Ang II had no additional effect, whereas it increased QO₂ in wild-type. Addition of H₂O₂ reduced transport-dependent QO₂ in wild-types, but not in p22(phox). Transport-independent QO₂ was unaffected by H₂O₂. The similar effects of Ang II and H₂O₂ to reduce transport-dependent QO₂ suggest a direct regulatory role of oxidative stress. In accordance, the transport-dependent QO₂ was reduced in p22(phox) already during baseline. The effects of Ang II on transport-independent QO₂ was not replicated by H₂O₂, indicating direct regulation via Ang II-receptors independently of oxidative stress. However, the Ang II effect was absent in p22(phox), suggesting that oxidative stress also modulates normal Ang II signaling. In conclusion, Ang II affects both transport-dependent and transport-independent QO₂ in proximal tubular cells and may be an important pathway modulating renal QO₂.


Beyond sequencing: machine learning algorithms extract biology hidden in Nanopore signal data.
Nanopore sequencing provides signal data corresponding to the nucleotide motifs sequenced. Through machine learning-based methods, these signals are translated into long-read sequences that overcome the read size limit of short-read sequencing. However, analyzing the raw nanopore signal data provides many more opportunities beyond just sequencing genomes and transcriptomes: algorithms that use machine learning approaches to extract biological information from these signals allow the detection of DNA and RNA modifications, the estimation of poly(A) tail length, and the prediction of RNA secondary structures. In this review, we discuss how developments in machine learning methodologies contributed to more accurate basecalling and lower error rates, and how these methods enable new biological discoveries. We argue that direct nanopore sequencing of DNA and RNA provides a new dimensionality for genomics experiments and highlight challenges and future directions for computational approaches to extract the additional information provided by nanopore signal data.


Normal mitotic reactivity of fibroblasts and mesothelial cells in thrombocytopenic rat.
Mitotic reactivity following 48/80-induced mast-cell secretion was studied in the mesentery of rats made thrombocytopenic, 7 days following a single injection of melphalan. In spite of a low platelet count (7% of normal), the mitogenic reaction of the mesenteric fibroblasts and mesothelial cells was normal as judged by DNA-synthesis and mitotic index. The findings suggest that platelets and platelet-growth factors are not essential for the mast-cell-mediated mitogenic reaction of these two types of connective-tissue cells studied in vivo.


Activation of complement by human hemoglobin and by mixtures of hemoglobin and bacterial endotoxin.
Purified human hemoglobin is being developed as an alternative to transfusions of homologous erythrocytes. However, toxicity associated with infusion of hemoglobin has limited the development of this resuscitation fluid. Some observed toxicities, including activation of the complement cascade, have been associated with contamination of hemoglobin solutions by bacterial endotoxin. Recent studies have demonstrated complex formation between hemoglobin and endotoxin, and have documented a resultant increase in the ability of endotoxin to activate coagulation, stimulate tissue factor production by human peripheral blood mononuclear cells, and stimulate tissue factor activity and protein synthesis in cultured human endothelial cells. The process of hemoglobin enhancement of endotoxin toxicity suggests a possible mechanism by which the consequences of endotoxin contamination of hemoglobin solutions, including complement activation, could be magnified. Therefore, we studied the potential of hemoglobin to either fix complement directly, or modify the ability of endotoxin to fix complement. Human crosslinked and native hemoglobins, at concentrations between 0.2 mg/ml and 3 mg/ml, were shown to fix complement. Complement fixation by hemoglobin was identical in normal human serum or in factor B-depleted serum, suggesting that fixation occurred via the classical pathway of complement activation. Complement fixation then was examined with a battery of smooth and rough endotoxins tested in the absence and presence of hemoglobin. Addition of hemoglobin to a solution of a rough Salmonella endotoxin partial structure, from which a single fatty acid had been hydrolyzed from the lipid A portion of the macromolecule, resulted in decreased efficiency of complement fixation. However, addition of hemoglobin had little or no effect on the intrinsic complement fixing abilities of eight other smooth endotoxins, rough endotoxins, or endotoxin partial structures. Our results demonstrated the ability of hemoglobin to fix complement at hemoglobin concentrations which would be achieved during infusion for resuscitation, but failed to demonstrate a reproducible effect of hemoglobin on the activation of complement by endotoxin.


Impaired motor learning attributed to altered AMPA receptor function in the cerebellum of rats with temporal lobe epilepsy: ameliorating effects of Withania somnifera and withanolide A.
The aim of this study was to investigate the effect of Withania somnifera (WS) extract, withanolide A (WA), and carbamazepine (CBZ) on cerebellar AMPA receptor function in pilocarpine-induced temporal lobe epilepsy (TLE). In the present study, motor learning deficit was studied by rotarod test, grid walk test, and narrow beam test. Motor learning was significantly impaired in rats with epilepsy. The treatment with WS and WA significantly reversed the motor learning deficit in rats with epilepsy when compared with control rats. There was an increase in glutamate content and IP3 content observed in rats with epilepsy which was reversed in WS- and WA-treated rats with epilepsy. alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor dysfunction was analyzed using radiolabeled AMPA receptor binding assay, AMPA receptor mRNA expression, and immunohistochemistry using anti-AMPA receptor antibody. Our results suggest that there was a decrease in Bmax, mRNA expression, and AMPA receptor expression indicating AMPA receptor dysfunction, which is suggested to have contributed to the motor learning deficit observed in rats with epilepsy. Moreover, treatment with WS and WA resulted in physiological expression of AMPA receptors. There was also alteration in GAD and GLAST expression which supplemented the increase in extracellular glutamate. The treatment with WS and WA reversed the GAD and GLAST expression. These findings suggest that WS and WA regulate AMPA receptor function in the cerebellum of rats with TLE, which has therapeutic application in epilepsy.


Breastfeeding the infant with congenital diaphragmatic hernia post extracorporeal membrane oxygenation.
Infants born with congenital diaphragmatic hernia (CDH) often require extracorporeal membrane oxygenation (ECMO). Infants on ECMO may experience a long period of being nothing by mouth (NPO) while receiving parenteral nutrition. Once the infant with CDH is repaired and off ECMO, human milk should be used to initiate enteral feedings. Human milk provides immunologic, developmental, and nutritional protection for these highrisk infants and may be crucial in decreasing morbidities commonly associated with post-ECMO survivors. These mother-infant dyads require extensive lactation support to ensure maintenance of milk supply and successful transition to direct breastfeeding. Three case studies are presented as exemplars to demonstrate how breastfeeding success can be achieved even in the most vulnerable infants.


Correction to: Sphenoid dysplasia in neurofibromatosis type 1: a new technique for repair.
The published version of this article unfortunately contained an error. All names of the authors have been published incorrectly. Given in this article are the corrected author names.


An increase of circulating leptin in patients with liver cirrhosis.
Leptin, the ob gene product, is an anorexigenic peptide secreted from adipose tissue. However, the mechanism of leptin clearance/degradation has not been well determined in humans. The present study was undertaken to examine a possible involvement of liver in determining circulating leptin concentrations in humans. In the present study 58 healthy control subjects and 68 patients with liver cirrhosis (LC) without any renal dysfunction were randomly included. The serum immunoreactive leptin (IRL) concentrations relative to the body mass index (BMI) were determined. Serum IRL and estradiol (E2) concentrations were assayed by radioimmunoassay (RIA). The correlations between the BMI and circulating IRL concentrations were all significant in male healthy controls (M-C), male patients with LC (M-LC), female healthy controls (F-C) and female patients with LC (F-LC). Circulating IRL concentrations were significantly higher than control in F-LC but not M-LC groups. The ratio of circulating IRL concentrations to the BMI was significantly higher in the M-LC group than in the M-C group and also significantly higher in the F-LC group than in the F-C group. The correlation between the IRL/BMI ratio and serum total bilirubin concentrations was significant (r = 0.417, P < 0.05) in the M-LC group, but not in the F-LC group. There was no significant correlation of the IRL/BMI ratio to serum E2 or albumin concentrations in either M-LC or F-LC groups. The present data demonstrated that the rate of increase in circulating IRL concentrations with the BMI was higher in LC patients of both genders. Liver may play a role in determining circulating leptin levels.


Effects of sympathetic innervation and temperature on the properties of rat heart adrenoceptors.
1. The pharmacological characteristics of adrenoceptors at different temperatures were assessed on the basis of the effects of various alpha- and beta-adrenoceptor agonists and antagonists on electrically-driven left atria and spontaneously-beating pairs of atria from rats. 2. Phenoxybenzamine (Pbz) potentiated inotropic responses of left atria to noradrenaline (NA) at 31 degrees C, produced significantly less potentiation at 24 degrees C and inhibited responses at 17 degrees C; it had little effect on responses to CaCl2. Both Pbz and phentolamine inhibited responses to phenylephrine more effectively at 17 than at 31 degrees C. N-cyclohexylmethyl-N-ethyl-beta-chloroethylamine hydrochloride (GD-131), a haloalkylamine with negligible alpha-adrenoceptor blocking activity, caused only potentiation of responses to NA at 17 degrees C. 3. The presence of phentolamine during incubation with Pbz eliminated block of responses to NA and revealed a potentiation that was equivalent at all three temperatures tested. Phentolamine did not alter the block of responses to 5-hydroxytryptamine by Pbz. Protection of alpha-adrenoceptors by phentolamine during exposure to [3H]-Pbz significantly decreased the amount of label bound to the myocardium at 17 degrees C, but did not alter binding at 31 degrees C. 4. Inhibition of responses to NA by propranolol decreased with temperature, and the magnitude of the change increased with the concentration of propranolol. Compared to 31 degrees C, the effect of the highest concentration of propranolol. (4.0 micronM) was significantly decreased at 24 degrees C, and the effects of all except the lowest concentration (0.04 micronM) were significantly decreased at 17 degrees C. 5. The potency of isoprenaline decreased and that of phenylephrine increased at low temperatures, and their potency ratio was much lower at 17 than at 31 degrees C for both the inotropic and chronotropic responses of spontaneously-beating atria. However, the ratio was unaffected by temperature in electrically-driven left atria. A similar difference between spontaneously-beating and driven preparations is apparent in the data of other workers, but its basis is not clear. 6. Atria from rats pretreated with 6-hydroxydopamine (6-OHDA) were sensitized to the effects of NA, and there was no increase in alpha-adrenoceptor properties at low temperatures. Little alpha-adrenoceptor activity could be demonstrated in chemically denervated atria at any temperature, 6-OHDA pretreatment did not alter the binding of [3H]-Pbz at 31 degrees C, but decreased it significantly at 17 degrees C. Pretreatment with reserpine caused some sensitization, but not significantly after the characteristics of the adrenoceptors or their responses to temperature. 7. It is concluded that the adrenoceptors of atria are affected by temperature in much the same way as those of frog hearts, although the transition from beta- to alpha-adrenoceptor properties may begin at a slightly higher temperature...


[Analysis of a mistake occurring during prenatal diagnosis of two couples respectively carrying CD41-42 (-TCTT) and CD43(G>T) mutations of the beta hemoglobin gene].
To provide prenatal diagnosis for two couples who respectively carried heterozygous CD41-42 (-TCTT) and CD43 (G>T) mutations of the beta hemoglobin gene. The mutations were simultaneously detected with reverse dot blot (two diagnostic kits), multi-color melting curve analysis and sequencing analysis. The fetus of family 1 was shown to be heterozygous for CD43 (G>T) by the three methods, while the fetus of family 2 was shown to be double heterozygous for CD41-42 (-TCTT) and CD43 (G>T) by multi-color melting curve analysis and sequencing analysis. The two diagnostic kits yielded different results by reverse dot blot, one as double heterozygous for CD41-42 (-TCTT) and CD43 (G>T), and another as homozygous for CD41-42 (-TCTT). For prenatal diagnosis of couples carrying mutations of beta hemoglobin gene such as CD41-42 (-TCTT) and CD43 (G>T), other methods such as Sanger sequencing should be used in order to avoid misdiagnosis.


The local effect of intravesical Corynebacterium parvum in the dog. A preliminary study.
High doses of C. parvum introduced in the bladder of the dog provoke changes in the submucosa. The significance and importance of these modifications should be investigated.