Dissolution enhancement of sorafenib tosylate by co-milling with tetradecanol post-extracted using supercritical carbon dioxide.

Sorafenib (SOR) is an important multikinase inhibitor for the treatment of cancers. It is commercially available (Nexavar from Bayer) in the form of sorafenib tosylate (SORt) due to its very low solubility. Studies have been made to further improve the dissolution behavior of the tosylate form (SORt), which could ultimately moderate the currently high daily dose. In the present study, SORt nanoparticles (SORt-NP) were prepared through a process that combined two industrially well-accepted techniques of co-milling and supercritical extraction. SORt was co-milled with hydrophilic polymers and tetradecanol, and the tetradecanol was post-extracted using supercritical carbon dioxide. The process enabled the formation of SORt-NP without using any toxic organic solvents, and the drug/excipient ratio (1:0.38) was substantially higher than determined in other studies (1:5.4-10). The enhanced dissolution behavior of SORt-NP was possible with an optimized number of milling cycles. Combining co-milling and supercritical extraction was able to form overall porous network structures with reduced crystallite size, which accelerated the dissolution of SORt-NP. The current method could be easily extended to other poorly soluble drugs as a general approach to improve their dissolution behaviors.

Assessment of the contact toxicity of methyl palmitate on Tetranychus viennensis (Acari: Tetranychidae).

Previous work demonstrated that methyl palmitate possesses acaricidal activity against Tetranychus viennensis Boisduval (Acari: Tetranychidae) via an unknown mechanism. Here, the symptoms of methyl palmitate toxicity to T. viennensi were studied to determine the acaricidal mechanism of action of this fatty acid methyl ester. Methyl palmitate caused concentration-dependent mortality of T. viennensis, with a moderate concentration (5 mg/ml) eliciting excitement and premature oviposition without spinning shortly after exposure. Tremors of the appendages were subsequently observed, followed by quiescence after approximately 5 h. Mites developed dorsal fluid exosmosis at 15-20 h posttreatment with reduced egg production, followed shortly thereafter by death. Some typical neurotoxic symptoms such as excitement and convulsions were observed in methyl palmitate-exposed mites, suggesting that methyl palmitate may be a neurotoxin. Compared with other neurotoxic acaricides, methyl palmitate poisoning is a slow process in mites. Transmission electron microscopy revealed serious ultrastructural damage in response to 5 mg/ml methyl palmitate exposure. Autolysis of membranous structures was also observed, especially in the mitochondria, suggesting a novel mode of action for methyl palmitate-induced toxicity.

The earliest human occupation of the high-altitude Tibetan Plateau 40 thousand to 30 thousand years ago.

The Tibetan Plateau is the highest and one of the most demanding environments ever inhabited by humans. We investigated the timing and mechanisms of its initial colonization at the Nwya Devu site, located nearly 4600 meters above sea level. This site, dating from 40,000 to 30,000 years ago, is the highest Paleolithic archaeological site yet identified globally. Nwya Devu has yielded an abundant blade tool assemblage, indicating hitherto-unknown capacities for the survival of modern humans who camped in this environment. This site deepens the history of the peopling of the "roof of the world" and the antiquity of human high-altitude occupations more generally.

LncRNA profiling of skeletal muscles in Large White pigs and Mashen pigs during development.

Long noncoding RNA (lncRNA) has been increasingly implicated in the regulation of muscle development. Large White pigs have a higher muscle growth rate than do Mashen pigs. In the present study, the lncRNA expression profiles in skeletal muscle of these 2 pig breeds were compared at 1, 90, and 180 d of age using RNA sequencing. We obtained 2,718 million clean reads and identified a total of 5,153 novel lncRNA. We found 1,407 differentially expressed lncRNA that showed consistent expression patterns between the 2 breeds at all the 3 sampling points. Ten lncRNA were randomly selected, and their expression was validated using Real-time Quantitative PCR. In summary, this study identifies a number of lncRNA that correlate with muscle growth. The regulation and function of these lncRNA in muscle growth and development need to be further explored.

Zygotic expression and activity of the Drosophila Toll gene, a gene required maternally for embryonic dorsal-ventral pattern formation.

Maternal expression of the Toll gene is required for the production and the correct spatial organization of all lateral and ventral structures of the Drosophila embryo. We show here that the Toll gene is transcribed zygotically in the embryo and that zygotic expression is important for the viability of the larva. Both genetic and molecular data indicate that the zygotic Toll product has the same biochemical activity as the maternal product. The spatial distribution of the Toll transcript in the embryo was analyzed. In contrast to the uniform distribution of the maternal RNA, the zygotic Toll RNA is present in a complex spatial and temporal pattern in the embryo. A striking feature of this pattern is the correlation of the regions of invaginating cells with sites of accumulation of zygotic Toll RNA.

Improvement in exercise capacity and exercise blood pressure response after transcoronary alcohol ablation therapy of septal hypertrophy in hypertrophic cardiomyopathy.

Transcoronary alcohol ablation (TAA) therapy of septal hypertrophy was recently proposed as a therapeutic modality for obstructive hypertrophic cardiomyopathy (HC). However, questions remain about the effect of TAA on exercise performance. We performed a time-course analysis of exercise capacity and exercise hemodynamics in 20 patients with symptomatic obstructive HC after TAA. Symptom-limited bicycle exercise testing was performed before and 3 and 12 months after TAA, and cardiac catheterization at 3-month follow-up. The pressure gradient of the left ventricular outflow tract immediately decreased from 58 +/- 8 to 4 +/- 1 mm Hg at rest (p <0.01) and from 143 +/- 11 to 30 +/- 6 mm Hg after extrasystole (p <0.01), but partially recovered at 3-month follow-up (14 +/- 4 and 40 +/- 9 mm Hg, respectively). Left ventricular end-diastolic pressure was not changed after TAA. Peak oxygen consumption increased from 19 +/- 2 to 23 +/- 1 ml/kg/min (p < 0.01) and exercise duration from 573 +/- 47 to 742 +/- 46 seconds (p <0.01) at 3-month follow-up, but thereafter reached a plateau. Abnormal patterns of exercise blood pressure response were shown in 9 patients but normalized after TAA. Major complications occurred in 4 patients: no reflow to the left anterior descending coronary artery in 2 patients and ventricular tachycardia requiring cardioversion in 2 patients. During the follow-up period, all patients survived with symptomatic improvement in 17 patients. Thus, TAA is a promising therapeutic modality with improvement in exercise capacity and abnormal exercise blood pressure response in obstructive HC. However, potential serious complications should be considered in the application of TAA.

Chromosome analysis of a newly established renal carcinoma cell line.

A renal cell carcinoma has been established as a cell line in vitro. Repeated chromosome analyses of the cell line revealed a stable clone with the modal chromosome number 80 and three pairs of marker chromosomes, M1-M3. M1 and M2 resulted from a translocation between a chromosome #3 deleted in band p14 and a normal #7: M1 = der(3)t(3;7) (:3p14----cen----3q24::7q21----7qter), and M2 = der(7)t(3;7)(3qter----3q24::7q21----cen----7pter ). M3 was a small metacentric chromosome, probably consisting of the centromeric portion of a #3: del(3)(:p14----cen----q12:). No other structural changes were present. Our findings are in agreement with those of previous studies, stating that rearrangements of 3p12-14 are primary cytogenetic events in renal cell carcinomas, even though this can only be inferred in this case. Thus, this cell line may be useful for further molecular and biochemical studies.

Isochromosomes i(8q) or i(9q) in three adenocarcinomas of the lung.

We have cytogenetically analyzed three primary adenocarcinomas of the lung. All tumors had chromosome numbers in the triploid region. The multiple structural aberrations included rearrangements of 3p, in two cases affecting the segment 3p14-23, where deletions are characteristically found in small cell lung carcinomas. Isochromosomes for 8q were present in two tumors and i(9q) in one tumor. In the few previously reported cytogenetic analyses of pulmonary adenocarcinomas, all of which examined metastases or cell lines, i(8q) was found in one case and i(9q) in two cases. These isochromosomes, therefore, represent previously unrecognized nonrandom changes in adenocarcinomas of the lung, and might constitute primary aberrations in this tumor type.

Simple numerical chromosome aberrations in well-differentiated malignant epithelial tumors.

Cytogenetic analysis of four well-differentiated malignant epithelial tumors revealed primary clones with only numerical abnormalities. The karyotypes were 49,XX, +5, +5, +7, +7, -17/50,XX, +5, +5, +7, +7, -17, +r in an adenocarcinoma of the lung; 47,XX, +3/47,XX, +5/47,XX, +7 in a squamous cell carcinoma of the epiglottis; 47,XX, +5/48,XX, +5, +10 in a squamous cell carcinoma developing in an ovarian dermoid cyst; and 52,XX, +5, +7, +8, +14, +15, +21 in a seropapillary ovarian adenocarcinoma. Also, in previously published cases exclusively numerical aberrations were much more common in highly differentiated epithelial tumors (22/74) than in moderately to low-differentiated carcinomas (13/281). Our findings and the literature data thus agree with a developmental scheme in which numerical changes, possibly reflecting an early-onset genomic instability in the tumor cells, may precede massive structural anomalies in the gradual malignization of epithelial tumors.

Two unrelated clonal chromosome rearrangements in a nasal papilloma.

We have cytogenetically examined short-term cultures from a nasal papilloma, a tumor type in which chromosome aberrations have hitherto not been reported. Two pseudodiploid clones were detected, giving the tumor karyotype 46,XY,t(1;3)(p31;p12)/46,XY,t(11;?)(q25;?).

Multiple apparently unrelated clonal chromosome abnormalities in a squamous cell carcinoma of the tongue.

We have cytogenetically examined short-term cultures from a squamous cell carcinoma of the tongue, a tumor type in which chromosome aberrations hitherto have not been reported. No less than 12 pseudodiploid clones were detected, giving the tumor karyotype 46,X,der(X)t(X;1)(q26;p32),der(1)(Xqter----Xq26::1p32 ----cen----1q42:), del(13)(q11q21),t(15;?) (q26;?)/46,XX,t(1;?)(p34;?),inv(2)(p21q11)/46,XX,t(1;10)(p32;q24)/ 46,XX, + der(1)(12pter----12p11::1p11----cen----1q32:: 11q13----11q22::1q32----1q42:), del(11)(q13q22), -12, der(17)t(1;17) (q42;p13)/46,XX,inv(1)(p22q44)/47,XX,del(1)(q32),der(17)t(1; 17)(p22;q25), der(1)inv(1) (q25q44)t(1;17)(p22;q25),ins(14;7)(q11;q22q36), + 14/46,XX,t(1;4)(q23;q35)/46, XX,t(1;21) (q25;q22),t(2;10)(q31;q26),t(22;?)(q12;?)/46,XX,del(1)(q32)/46,XX, t(1;8)(q44;q21)/46,XX, t(2;21)(q11;p11)/46,XX,t(9;11)(q34;q13). The large number of apparently unrelated abnormalities leads us to suggest that the carcinoma may have been of multiclonal origin.

Multiple clonal chromosome aberrations in squamous cell carcinomas of the larynx.

Short-term cultures from five squamous cell carcinomas of the larynx were subjected to cytogenetic analysis. In the first three cases, two, three, and 10 chromosomally abnormal clones were detected. Single clonal abnormalities were found in cases 4 and 5. In addition to the clonal aberrations, a number of nonclonal changes were also present in all five tumors. None of the aberrations, clonal or nonclonal, was found in more than one tumor, nor did the rearrangements correspond to any of the consistently cancer-associated aberrations known from other tumors. The remarkably diverse karyotypic picture of the five squamous cell larynx carcinomas, in particular the finding of cytogenetically unrelated clones in three of them, suggests that some of these neoplasms are polyclonal rather than monoclonal.

Chromosome rearrangements in two uterine sarcomas.

Cytogenetic analysis of short-term cultures from two uterine sarcomas revealed clonal chromosome abnormalities in both cases. A locally recurrent mixed mesodermal tumor had the karyotype 61,XX,+2,+3,+del(5)(q11),+6,+7,+del(7)(q32),+8,+8,+8,+10, -11,-11,+der(11)t(1;11)(q12;p15),+der(11)t(1;11)(q12;p15),+der(11)t(1;11)(q12;p15),+del(12)(q14q21),+13,+15,del(17)(q23),+20. The other tumor, a lung metastasis from a uterine leiomyosarcoma, had several karyotypically abnormal clones. Two of them consisted of highly aberrant cells with modal chromosome numbers of 82 and 153, respectively, but because of insufficient quality the complex anomalies could not be identified. Various chromosomal changes that included translocations, deletions, insertions, and numerical rearrangements (always with extra chromosome 7 material) were identified in pseudo- or near-diploid cells, resulting in nine additional cytogenetically abnormal clones.

Unique karyotypic abnormalities in a squamous cell carcinoma of the larynx.

We have cytogenetically examined short-term cultures from a squamous cell carcinoma of the larynx, a type of carcinoma in which chromosome aberrations have hitherto not been reported. The tumor karyotype was 46,XY,inv(2)(p22q24),t(9;13)(q34;q12),t(11;18)(q23;q21). None of these abnormalities have been described in carcinomas before.

Diverse chromosome abnormalities in squamous cell carcinomas of the skin.

Short-term cultures from three invasive squamous cell carcinomas of the skin were cytogenetically analyzed. Clonal chromosome aberrations were found in all tumors. In the first case, two of three abnormal clones were related, and in the second case, two of five clones demonstrated cytogenetic similarities. Both clones detected in case 3 had a structural rearrangement in common. Several nonclonal changes were seen in all three cases in addition to the clonal aberrations. None of the rearrangements detected, clonal or nonclonal, corresponds to any of the consistently cancer-associated aberrations known from other neoplasms. The remarkably diverse karyotypic picture of the three squamous cell carcinomas, in particular the finding of unrelated clones in two of them, hints that these neoplasms may be poly-rather than monoclonal. The lack of a common cytogenetic denominator argues that if chromosomal changes are of pathogenetic importance in this tumor type, a wide variety of apparently dissimilar changes exist that are roughly equal in their capacity to malignantly transform skin epithelium.

Basosquamous papilloma. A benign epithelial skin tumor with multiple cytogenetic clones.

Cytogenetic analysis of short-term cultures from a basosquamous papilloma revealed the following mosaic karyotype: 46,XX,t(2;5)(q31;q31),t(8;15)(p21;q21)/46,XX,t(7;17)(p13;p13)/47,XX, t(3;20)(q12;p13),+7/46,XX,t(1:12)(p12;q13). The finding of four abnormal, cytogenetically unrelated clones suggests a multicellular origin of this benign skin tumor. None of the structural rearrangements encountered have previously been associated with neoplasia.

Ethanol and thermotolerance in the bioconversion of xylose by yeasts.

The mechanisms underlying ethanol and heat tolerance are complex. Many different genes are involved, and the exact basis is not fully understood. The integrity of cytoplasmic and mitochondrial membranes is critical to maintain proton gradients for metabolic energy and nutrient uptake. Heat and ethanol stress adversely affect membrane integrity. These factors are particularly detrimental to xylose-fermenting yeasts because they require oxygen for biosynthesis of essential cell membrane and nucleic acid constituents, and they depend on respiration for the generation of ATP. Physiological responses to ethanol and heat shock have been studied most extensively in S. cerevisiae. However, comparative biochemical studies with other organisms suggest that similar mechanisms will be important in xylose-fermenting yeasts. The composition of a cell's membrane lipids shifts with temperature, ethanol concentration, and stage of cultivation. Levels of unsaturated fatty acids and ergosterol increase in response to temperature and ethanol stress. Inositol is involved in phospholipid biosynthesis, and it can increase ethanol tolerance when provided as a supplement. Membrane integrity determines the cell's ability to maintain proton gradients for nutrient uptake. Plasma membrane ATPase generates the proton gradient, and the biochemical characteristics of this enzyme contribute to ethanol tolerance. Organisms with higher ethanol tolerance have ATPase activities with low pH optima and high affinity for ATP. Likewise, organisms with ATPase activities that resist ethanol inhibition also function better at high ethanol concentrations. ATPase consumes a significant fraction of the total cellular ATP, and under stress conditions when membrane gradients are compromised the activity of ATPase is regulated. In xylose-fermenting yeasts, the carbon source used for growth affects both ATPase activity and ethanol tolerance. Cells can adapt to heat and ethanol stress by synthesizing trehalose and heat-shock proteins, which stabilize and repair denatured proteins. The capacity of cells to produce trehalose and induce HSPs correlate with their thermotolerance. Both heat and ethanol increase the frequency of petite mutations and kill cells. This might be attributable to membrane effects, but it could also arise from oxidative damage. Cytoplasmic and mitochondrial superoxide dismutases can destroy oxidative radicals and thereby maintain cell viability. Improved knowledge of the mechanisms underlying ethanol and thermotolerance in S. cerevisiae should enable the genetic engineering of these traits in xylose-fermenting yeasts.

Brain potentials related to seeing one's own name.

Subjects were assigned an assumed name and then shown a series of statements of the form, "My name / is / X", where X was the assumed name, their own first name, or one of a set of other false names. Their task was to respond positively to the "assumed" name and reject as false all other names, including their own. An N380 feature of the averaged task-related brain potentials, considered to be inversely related to the degree of contextual priming, was greatly enhanced for the false names compared to the assumed name. The N380 to one's own name was more similar to that of the false than the assumed name, indicating that the sentence context's priming of various names was under the subjects' attentional control, and that the late negativity could be modulated by this attention. In contrast, a large P510 feature distinguished one's own name from the false name, and this difference was unaffected by practice. Even in cases, then, where the context allows anticipation of one verbal event (here, the assumed name), a highly overlearned and salient stimulus such as one's own name continues to produce a distinctive neural response.

Expression of erythropoietin and its receptor in kidneys from normal and cyclosporine-treated rats.

Long-term treatment with cyclosporine A (CsA) is associated with various types of complications; however, CsA-induced anemia has not been reported. The present study examined the impact of CsA on hematopoietic parameters and intrarenal expression of erythropoietin (EPO) and the EPO receptor (EPOR) in a rat model of chronic CsA nephrotoxicity. Sprague-Dawley rats were fed a low-salt diet (0.05% sodium) and were treated daily for 4 weeks with vehicle (olive oil 1 mL/kg subcutaneously) or CsA (15 mg/kg subcutaneously). The expression of EPO and EPOR was evaluated by immunohistochemistry and immunoblotting, and hematopoietic parameters were assessed by measuring blood hemoglobin and hematocrit levels, and these variables were compared between treatment groups. Renal function, oxidative stress, histopathology (tubulointerstitial fibrosis), apoptotic cell death, and expression of transforming growth factor ß-inducible gene-h3 (ßig-h3) were also compared between treatment groups. In kidneys from vehicle-treated rats, endogenous EPO and EPOR protein were expressed constitutively in the outer stripe of the outer medulla and the cortex. EPO protein expression decreased significantly in kidneys from CsA-treated rats. By contrast, EPOR expression was higher in kidneys from CsA-treated rats than in vehicle-treated rats. These changes were accompanied by decreases in serum hemoglobin and hematocrit levels and correlated with the number of cells positive for terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (r = -0.769, P = .003) and ßig-h3 protein expression (r = -0.910, P < .001). Long-term treatment with CsA suppresses renal endogenous EPO expression, resulting in anemia. Increases in apoptotic cell death and ßig-h3 expression are closely associated with inhibition of EPO expression in chronic CsA nephrotoxicity.

High voltage ultrawide band pulse generator using Blumlein pulse forming line.

A high voltage ultrawide band pulse generation system has been developed to radiate intense and ultrawide band electric fields for the examination of effects of the electric fields on the operation of electronic devices. As major components of the system, a helical strip∕wire type of air-cored pulse transformer and a triaxial type of Blumlein pulse forming line have been designed and fabricated to amplify and shape the output pulse, respectively. For the construction of a compact system, the pulse transformer and the Blumlein line are installed in a single cylindrical container. An ultrawide band TEM horn antenna has been fabricated to radiate the Blumlein output pulses to electronic devices. A number of experimental results demonstrate that the system is capable of providing an output pulse whose voltage is greater than 300 kV, pulse duration is ~5 ns, and rise time is ~500 ps with repetition rate of 10 Hz. The peak-to-peak value of electric field intensity of a radiated pulse is also measured to be approximately 42 kV/m at a distance of 10 m away from the antenna.