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.

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.

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.

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.

Metabolic engineering for improved fermentation of pentoses by yeasts.

The fermentation of xylose is essential for the bioconversion of lignocellulose to fuels and chemicals, but wild-type strains of Saccharomyces cerevisiae do not metabolize xylose, so researchers have engineered xylose metabolism in this yeast. Glucose transporters mediate xylose uptake, but no transporter specific for xylose has yet been identified. Over-expressing genes for aldose (xylose) reductase, xylitol dehydrogenase and moderate levels of xylulokinase enable xylose assimilation and fermentation, but a balanced supply of NAD(P) and NAD(P)H must be maintained to avoid xylitol production. Reducing production of NADPH by blocking the oxidative pentose phosphate cycle can reduce xylitol formation, but this occurs at the expense of xylose assimilation. Respiration is critical for growth on xylose by both native xylose-fermenting yeasts and recombinant S, cerevisiae. Anaerobic growth by recombinant mutants has been reported. Reducing the respiration capacity of xylose-metabolizing yeasts increases ethanol production. Recently, two routes for arabinose metabolism have been engineered in S. cerevisiae and adapted strains of Pichia stipitis have been shown to ferment hydrolysates with ethanol yields of 0.45 g g(-1) sugar consumed, so commercialization seems feasible for some applications.

Maturation of xenoantibody gene expression during the humoral immune response of rats to hamster xenografts.

Immunoglobulin isotype switching represents an important component of antibody maturation in the development of humoral immune responses. We have recently conducted a series of studies in a nonimmunosuppressed rodent model to define the kinetics of xenoantibody production and seek evidence for the maturation of xenoantibody Ig gene expression by xenograft recipients. LEW rats were transplanted with hamster cardiac xenografts and the grafts were allowed to remain in situ for prolonged immune stimulation of the host. Anti-hamster antibodies were examined at days 4, 8, 21, 28 and 40 post-transplantation. cDNA libraries specific for rat mu or gamma heavy chains were constructed from B lymphocytes of the xenograft recipients at day 4 and day 21 post-transplantation. Selected cDNA clones encoding the Ig V(H)HAR family of genes from each group were sequenced and analyzed for the presence of somatic mutations. We found that the reactivity of xenoantibodies examined with flow cytometry underwent sequential changes in which IgM titers peaked at day 8 post-transplantation (PTx) and returned to low levels after 21 days. IgG titers started to increase at about one week PTx and peaked at 21-28 days. All the IgG isotypes (IgG1, 2a, 2b and 2c) were differentially involved in the IgG responses. Serum passive transfer experiments demonstrated that IgM antibody fractions separated from sera at day 4 post-transplantation were capable of causing hyperacute rejection (HAR) of hamster xenografts, whereas IgM fractions from days 21-40 failed to cause HAR (N = 7, MST = 4 days), a pattern that was consistent with a rise in total xenoreactive IgM levels at days 4-8 and a fall to low levels at 21 days post-transplantation. IgG-containing fractions separated from day 21-40 antisera caused HAR (N = 7, MST = 36 min) whereas IgG fractions from day 8 sera failed to induce graft rejection. Genetic analysis of the rearranged VH genes from 10 cDNA clones demonstrated that the Ig mu (n = 5) and gamma (n = 5) chain clones used the same family of VH genes (V(H)HAR family) to encode their antibody binding activity. The majority (80%) of the IgM clones were present in their original germline configuration. In contrast, the nucleotide sequences from IgG clones manifested an increase in the numbers of replacement mutations in the CDR region of the Ig heavy chain genes, providing evidence for a potential role for somatic mutation in the maturation of IgG xenoantibody responses as the humoral response matures with time post-transplantation.

Korean beliefs about everyday memory and aging for self and others.

Studies in the West have demonstrated that more everyday memory problems are expected for typical older adults than for typical young adults. In order to examine memory beliefs about aging in Asia, we conducted a study in Korea which parallels that of Ryan and Kwong See (1993). We used the three self-efficacy scales of the Metamemory in Adulthood instrument (Dixon & Hultsch, 1983) to determine whether age changes are anticipated for oneself as well as for typical adults. Young adults (N = 468; mean age = 21.0 years) rated the memory of either typical adults (aged 25, 45, or 65 years) or themselves at one of these ages. As in Ryan and Kwong See (1993), anticipation of decline was obtained on two of the three self-efficacy scales (i.e., capacity and change). In addition, beliefs about everyday memory decline (i.e., capacity and locus) were weaker for the self than for typical others. Hence, support was obtained for negative stereotypes about memory and aging in Korea as well as a self-protection bias indicating stronger anticipation of age-related decline among others.

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.

Alterations of the actin polymerization status as an apoptotic morphological effector in HL-60 cells.

The alterations of the cytoskeletal actin network have been implicated as a morphological effector in apoptosis. However, studies directly linking actin change to the morphological events in apoptosis are lacking. This study quantitatively examined the effect of actin alteration on the camptothecin (CPT)-induced apoptotic process in HL-60 cells. Actin alteration was induced by two distinctive types of agent: the polymerization-stimulating agent, Jasplakinolide (Jas), and the polymerization-blocking agent, cytochalasin B (CB). The actin polymerization status was measured by two complementary methods: the cell pellet-based DNase I inhibition method, and the individual cell-based quantitative fluorescence image analysis (QFIA) assay. Actin polymerization induced by Jas caused apoptosis directly. By contrast, CB, an actin polymerization-blocking agent, partially inhibited CPT-induced apoptosis. A similar inhibition of the CPT-induced apoptosis response was observed with a more specific actin depolymerization agent, cytochalasin E. The alterations of the actin polymerization status occurred in three sequential steps during the apoptotic process: first polymerization, followed by depolymerization, and finally degradation. However, compared with CPT-induced apoptosis, Jas-induced apoptosis was characterized by pronounced actin polymerization that corresponded morphologically with prominent membrane blebbing, but less apoptotic body formation. Furthermore, DNase I activity, which is normally inhibited by G-actin, was specifically detected in Jas-treated cells. These results show that the regulation of actin polymerization is an important apoptotic morphological effector, whereas the alterations of the actin polymerization status by chemicals have profound effects not only on altering the morphology of apoptotic cells, but on apoptosis induction in HL-60 cells as well.

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.

Effects of percutaneous balloon mitral valvuloplasty and exercise training on the kinetics of recovery oxygen consumption after exercise in patients with mitral stenosis.

AIMS: Kinetics of recovery oxygen consumption after exercise plays an important role in determining exercise capacity. This study was performed to assess the kinetics of recovery oxygen consumption in mitral stenosis and evaluate the effects of percutaneous balloon mitral valvuloplasty and exercise training on the kinetics. METHODS AND RESULTS: Thirty patients with mitral stenosis (valve area < or =1.0 cm2) and same sized age- and size-matched healthy volunteers were included for this study. All subjects performed maximal upright graded bicycle exercise. Thirty consecutive patients who underwent successful percutaneous balloon mitral valvuloplasty (valve area > or =1.5 cm2 and mitral regurgitation grade < or =2), were randomized to an exercise training group or non-training group. The exercise group performed daily exercise training for 3 months. Half-recovery time of peak oxygen consumption was significantly delayed in mitral stenosis as compared to normal subjects (120+/-42 s vs 59+/-5, P<0.01). Peak oxygen consumption (ml x min(-1) x kg(-1)) was significantly increased in both the training (16.8+/-4.9 to 25.3+/-6.9) and non-training groups (16.3+/-5.1 to 19.6+/-6.0) 3 months after percutaneous balloon mitral valvuloplasty. Half-recovery time of peak oxygen consumption was significantly shortened in the training group (124+/-39 to 76+/-13, P<0.01), but not in the non-training group (114+/-46 to 109+/-44 s, P=0.12) at 3 months follow-up. The degrees of symptomatic improvement after percutaneous balloon mitral valvuloplasty were more closely correlated with the changes of the half-recovery time of peak oxygen consumption than those of peak oxygen consumption. CONCLUSION: Kinetics of recovery oxygen consumption was markedly delayed in mitral stenosis, which was improved after exercise training but not after percutaneous balloon mitral valvuloplasty alone. These results suggest that adjunctive exercise training may be useful for improvement of recovery kinetics and subjective symptoms after percutaneous balloon mitral valvuloplasty.

Effects of gamma irradiation on red cells from donors with sickle cell trait.

BACKGROUND: Transfusion-associated graft-versus-host disease can be prevented by gamma irradiation of blood components. Red cells (RBCs) from sickle cell disease patients may exhibit oxidative changes of RBC membranes due to the instability of hemoglobin (Hb) S. Persons with sickle cell trait are eligible to donate blood, and 35 to 45 percent of their total Hb is Hb S. The effect of gamma irradiation on RBCs from such persons is of interest. STUDY DESIGN AND METHODS: RBCs from 12 donors with sickle cell trait (Hb AS) and from 12 with normal Hb (Hb AA) were studied. Each of the 24 RBC units was divided equally into two transfer bags via a sterile connecting device. One bag from each RBC unit received a 2500-cGy dose of gamma irradiation at its mid-plane and was stored at 4 degrees C; the second set of bags was stored without irradiation. For RBCs from 6 donors with Hb AS and 6 donors with Hb AA, units were irradiated on Day 7 and studied on Day 35 of storage (Group 1). For the RBCs from the other 6 donors with Hb AS and the other 6 donors with Hb AA, units were irradiated on Day 28 and studied on Day 42 of storage (Group 2). RESULTS: For Group 1 and Group 2, plasma potassium and plasma Hb concentrations were significantly higher and RBC ATP concentrations were slightly lower in the irradiated units than in the nonirradiated units. In Group 1 and Group 2, there were no significant differences in the plasma potassium or RBC ATP concentrations in either the irradiated or the nonirradiated units of RBCs from donors with Hb AS and donors with Hb AA. Plasma Hb concentrations were consistently lower in the units from donors with Hb AS, whether or not they were irradiated. However, in both groups, proportionally similar changes in plasma Hb concentration were detected when the irradiated Hb AS and Hb AA units were compared to nonirradiated Hb AS and Hb AA units. CONCLUSION: Gamma irradiation of RBCs from donors with Hb AS or with Hb AA resulted in comparable changes in plasma potassium, RBC ATP, and plasma Hb concentrations, although donors with Hb AS had lower plasma Hb. RBCs from donors with Hb AS subjected to 2500 cGy of gamma irradiation did not evidence a storage lesion greater than that seen in RBCs from donors with Hb AA.

Comparison of platelet loss during leukocyte reduction at 4, 24, and 48 hours postcollection using a Closed System Apheresis Kit with Integral filter.

Prestorage leukocyte reduction of platelet concentrates may reduce adverse effects of transfusion while affording better quality control. Platelets and leukocytes may undergo activation during storage, which could affect the performance of leukocyte reduction filters. The purpose of this study was to evaluate the efficiency of leukocyte reduction and concomitant platelet loss with a new apheresis kit with an integral leukocyte reduction filter. Twelve donors underwent plateletpheresis on three occasions using the CS-3000 PLUS Blood Cell Separator with the Access Management System and the Access Closed System Apheresis Kit with Integral Sepacell Leukocyte Reduction Filter and Double Return Line Needle (Baxter-Fenwal Division, Deerfield, IL). Of the three products from each donor, one each was filtered at 4, 24, and 48 hours after completion of the plateletpheresis. Mean prefiltration platelet count was 4.43 x 10(11) and mean postfiltration platelet count was 3.56 x 10(11). Mean platelet recovery at 4, 24, and 48 hours filtration was 75%, 83%, and 84%, respectively. Analysis of variance (ANOVA) demonstrated that platelet recovery with filtration at four hours was significantly less than with filtration at 24 hours (P = 0.0236) and filtration at 48 hours (P = 0.0122). Platelet recovery with filtration at 24 hours did not differ significantly from filtration at 48 hours (P = 0.7684). Mean prefiltration WBC count was 0.93 x 10(6) and mean postfiltration WBC count was 0.12 x 10(6). Efficiency of leukocyte reduction was not significantly related to when filtration was performed. There was no significant variation from donor in platelet recovery or in leukocyte reduction efficiency. This method of prestorage leukocyte reduction demonstrated slightly but statistically significantly better platelet recovery with filtration at 24 or 48 hours after platelet collection compared to four hours. All filtration times provided acceptable platelet yields with very low residual WBC.

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.

Frequent rearrangement of chromosomal bands 1p22 and 11q13 in squamous cell carcinomas of the head and neck.

We report the finding of clonal structural chromosome abnormalities in short-term cultures from 15 squamous cell carcinomas of the head and neck region. When the distribution of chromosomal breakpoints in these 15 tumors and in the 16 head and neck carcinomas previously described are assessed, a marked clustering is seen at bands 1p22 and 11q13, which are rearranged in eight and nine tumors, respectively. No other band was involved in aberrations in more than five tumors. Cytogenetic evidence of gene amplification was seen in four tumors, three times in the form of homogeneously staining regions (twice located in 11q13), and in one tumor as double minutes. Among the candidate genes for such amplification are BCLI, INT2, and HSTI, all of which map to 11q13, and NRAS, which maps to 1p22. All these oncogenes have previously been shown to be amplified in subsets of head and neck carcinomas. We conclude that bands 1p22 and 11q13 are nonrandomly involved in chromosomal rearrangements in head and neck carcinomas and suggest that activation of oncogenes located in these bands may proceed via cytogenetic mechanisms.

Trisomy 5 and loss of the Y chromosome as the sole cytogenetic anomalies in a cavernous hemangioma/angiosarcoma.

Cytogenetic analysis of a cavernous hemangioma with transition to angiosarcoma revealed the mosaic karyotype 47,XY,+5/46,X,-Y,+5/45,X,-Y/46,XY. No cytogenetically analyzed hemangiomas or angiosarcomas have been reported before.

Dominant and recessive alleles of the Drosophila easter gene are point mutations at conserved sites in the serine protease catalytic domain.

The easter gene, required for the development of all lateral and ventral pattern elements in the Drosophila embryo, appears to encode an extracellular serine protease. Dominant easter alleles increase the number of cells that give rise to lateral and ventral structures. We have found that nine dominant and four recessive mutations are caused by single amino acid substitutions at conserved sites in the putative serine protease catalytic domain. The activity of dominant products was assayed by injecting in vitro synthesized transcripts from the dominant alleles into young embryos. The results suggest that the dominant easter products cleave the normal substrate, but fail to respond to a spatially asymmetric regulator.