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Several lines of research suggest that BCL-XL-mediated anti-apoptotic effects may contribute to pathogenesis of myeloproliferative neoplasms driven by JAK2V617F and serve as therapeutic target. Here, we used a knock-in JAK2V617F mouse model and confirmed that Bcl-xl was overexpressed in erythroid progenitors. MPN-induced phenotype in the peripheral blood by conditional knockin of JAK2V617F was abrogated by conditional knockout of Bcl2l1, which presented anemia and thrombocytopenia independently of JAK2 mutation status. The Mx1-Cre Jak2V617W/VF /Bcl2l1f/f mice presented persistent splenomegaly as a result of extramedullary hematopoiesis and pro-apoptotic stimuli in terminally differentiated erythroid progenitors. The pan-BH3 mimetic inhibitor obatoclax showed superior cytotoxicity in JAK2V617F cell models, and reduced clonogenic capacity in ex vivo assay using Vav-Cre Jak2V617F bone marrow cells. Both ruxolitinib and obatoclax significantly reduced spleen weights in a murine Jak2V617F MPN model but did not show additive effect. The tumor burden reduction was observed with either ruxolitinib or obatoclax in terminal differentiation stage neoplastic cells but not in myeloid-erythroid precursors. Therefore, disrupting BCL2 balance is not sufficient to treat MPN at the stem cell level, but it is certainly an additional option for controlling the critical myeloid expansion of the disease.

The induction of apoptosis in vivo is a useful tool for investigating the functions and importance of particular tissues. B-cell leukaemia/lymphoma 2-associated X protein (Bax) functions as a pro-apoptotic factor and induces apoptosis in several organisms. The Bax-mediated apoptotic system is widely conserved from Caenorhabditis elegans to humans. In order to establish a tissue-specific cell death system in the domestic silkworm, Bombyx mori, we constructed a transgenic silkworm that overexpressed mouse Bax (mBax) in particular tissues by the Gal4-upstream activation sequence system. We found that the expression of mBax induced specific cell death in the silk gland, fat body and sensory cells. Fragmentation of genomic DNA was observed in the fat body, which expressed mBax, thereby supporting apoptotic cell death in this tissue. Using this system, we also demonstrated that specific cell death in sensory cells attenuated the response to the sex pheromone bombykol. These results show that we successfully established a tissue-specific cell death system in vivo that enabled specific deficiencies in particular tissues. The inducible cell death system may provide useful means for industrial applications of the silkworm and possible utilization for other species.

SUMMARY Background: Despite improvements in first-line therapies, the outcomes of relapsed or refractory childhood acute leukaemia that has not achieved complete remission after relapse, has relapsed after stem cell transplantation (SCT), has primary induction failure and has relapsed with a very unfavourable cytogenetic risk profile, are dismal. Objectives and Methods: We evaluated the feasibility and efficacy of T-cell-replete haploidentical peripheral blood stem cell transplantation (haplo-SCT) with low-dose anti-human thymocyte immunoglobulin (ATG), tacrolimus, methotrexate and prednisolone (PSL) in 14 paediatric patients with high-risk childhood acute leukaemia. Results: All patients achieved complete engraftment. The median time to reaching an absolute neutrophil count of more than 0.5 × 10 9 L −1 was 14 days. Acute graft-vs-host disease (aGVHD) of grades II–IV and III–IV developed in 10 (71%) and 2 (14%) patients, respectively. Treatment-related mortality and relapse occurred in one (7%) patient and six (43%) patients, respectively. Eleven patients were alive and seven of them were disease-free with a median follow-up of 36 months (range: 30–159 months). The probability of event-free survival after 2 years was 50%. Conclusion: These findings indicate that T-cell-replete haplo-SCT, with low-dose ATG and PSL, provides sustained remission with an acceptable risk of GVHD in paediatric patients with advanced haematologic malignancies.

The patient was a 58-year-old man. A recent complete work-up was done to find the cause of epigastric pain and revealed a nodule about 4 cm in diameter in the upper right abdomen on CT scans. Laparoscopic resection was performed to allow for a definitive diagnosis to be made and to treat the lesion. The tumor was located in the transverse mesocolon, and there was no communication between the lesion and the ascending or transverse colon. Spindle-shaped tumor cells were arranged in palisades. The number of mitotic figures was only 1/50 HPF. The tumor was weakly positive for KIT and negative for CD34. From these findings, a diagnosis of extra-gastrointestinal stromal tumor originating in the transverse mesocolon was made.

Contents The purpose of this study was to assess the feasibility of polyester mesh culture for the in vitro production of bovine embryos, as polyester mesh is an alternative way for tracking individual embryos throughout culture using time-lapse cinematography (TLC). Bovine embryos were isolated during in vitro culture using sections of three different polyethylene terephthalate (PET) mesh products. In vitro matured and fertilized bovine oocytes were cultured in the 217 × 217, 230 × 230 or 238 × 238-μm openings of PET mesh sections or in simple micro-drops (control) for 7 days under either 20% or 5% O2 tensions. No difference in embryo developmental rates was found between the culture groups in terms of cleavage, blastocyst formation and blastocyst expansion irrespective of O2 tension. In contrast, under 20% O2 tension, blastocysts that developed in PET mesh with 217 × 217-μm opening had significantly higher numbers of total and trophectoderm (TE) cells than control embryos; however, the numbers and proportions of inner cell mass (ICM) cells did not differ. Under 5% O2 tension, no difference was found among the culture groups in the numbers of total, ICM and TE cells in embryos. All three PET mesh products investigated in this study were proven to be effective to prevent embryo movement. The results demonstrate that bovine embryos can be cultured in PET mesh sections without negative side-effects and suggest that embryo distance determined by the mesh affects embryo quality at atmospheric oxygen tension. Polyethylene terephthalate mesh with 217 × 217-μm openings was found to be the most suitable for further application in TLC.

Radiance measurements from the Stratospheric Sounding Unit (SSU) and the Advanced Microwave Sounding Unit (AMSU-A) are the primary source of information for stratospheric temperature in reanalyses of the satellite era. To improve the time consistency of the reanalyses, radiance biases need to be properly understood and accounted for in the assimilation system. The investigation of intersatellite differences between SSU and AMSU-A radiance observations shows that these differences are not accurately reproduced by the operational version of the radiative transfer model for the TIROS Operational Vertical Sounder (RTTOV-8). We found that this deficiency in RTTOV was mainly due to the treatment of the Zeeman effect (splitting of the oxygen absorption lines at 60 GHz) and to changes in the spectral response function of the SSU instrument that are not represented in RTTOV. On this basis we present a revised version of RTTOV that can reproduce SSU and AMSU-A intersatellite radiance differences more accurately. Assimilation experiments performed with the revised version of RTTOV in a four-dimensional variational analysis system (4D-Var) show some improvements in the stratospheric temperature analysis. However, significant jumps in the stratospheric temperature analysis still occur when switching satellites, which is due to the fact that systematic errors in the forecast model are only partially constrained by observations. Using a one-dimensional retrieval equation, we show that both the extent and vertical structure of the partial bias corrections must inevitably change when the nature of the radiance measurement changes with the transition from SSU to AMSU-A. Copyright © 2009 Royal Meteorological Society

Summary. A total of 143 cerebrospinal and serum samples, from proven and suspected cases of cryptococcosis, were concurrently examined using a recently introduced enzyme immunoassay (EIA Premier, Meridian Diagnostics, Inc., Cincinnati, OH, USA) and three latex agglutination (LA) procedures (Immunomycologics, Inc., Norman, OK, USA; IBL, Inc., Cranbury, NJ, USA and a non-commercial LA test). Of these 143 specimens, 115 were negative for cryptococcal antigen (CrAg) with the EIA and LA tests. The remaining 28 specimens were evaluated by the LA tests, and all were positive for CrAg (with titres ranging from 1:2 to 1:8192). Of these 28 LA-positive specimens, 26 were also tested by the EIA. This procedure detected CrAg in 23 specimens (88.5%), with antigen levels ranging from 1:4 to 1:266,857. There were 3 LA-positive specimens (titres 1:4 to 1:32) which were negative by the EIA procedure (10.7%). One LA-negative specimen demonstrated CrAg (titre 1:30) by the EIA procedure. The sensitivity of the EIA and LA tests was 85.2 and 100%, respectively. The specificity of the LA test was 100%, whereas that of the EIA was 97%. The agreement among laboratories for testing the specimens with the three LA tests was 100%. Zusammenfassung. An 143 Liquor- und Serumproben von Patienten mit bewiesener Cryptococcose oder Cryptococcose-Verdacht wurde ein neu eingefuhrter Enzymimmunoassay (EIA Premier, Meridian Diagnostics, Inc., Cincinnati, OH, USA) zum Cryptococcus-Antigen-nachweis (CrAG) vergleichend mit drei Latex-Agglutinationssystemen (LA; Immunomycologics, Inc., Norman, OK, USA; IBL, Inc., Cranbury, NJ, USA; und ein nicht-kommerzieller LA-Test) bewertet. Von den 143 Proben waren 115 CrAG-negativ im EIA und in den LA-Tests. Die ubrigen 28 Proben waren in den LA-Tests CrAG-positiv rnit Titern von 1:2 bis 1:8192. Von diesen 28 Proben wurden 26 auch im EIA gepruft. Dieses System reagierte bei 23 Proben (88.5%) CrAG-positiv mit Titern von 1:4 bis 1:266 857. Drei Proben, die in den LA-tests mit Titern von 1:4 bis 1:32 CrAG-positiv waren, reagierten im EIA CrAG-negativ (10.7%). Eine in den LA-Tests CrAG-negative Probe reagierte im EIA rnit einem Titer von 1:30 CrAG-positiv. Die Sensitivi-tat des EIA betrug 85.2%, die der LA-Tests 100%. Die Spezifitat der LA-Tests lag bei 100%, die des EIA bei 97%. Die Ubereinstimmung der Ergebnisse der drei LA-Tests zwischen den drei Testlaboratorien betrug 100%.

Avian influenza outbreaks caused by a low-pathogenic H5N2 virus occurred in Japan from June to December 2005. All 41 affected farms housed layer chickens. Therefore, we conducted a case-control study targeting all commercial layer chicken farms within the movement restriction areas in Ibaraki prefecture, where most outbreaks were detected, to investigate the risk factors for the introduction of avian influenza virus (AIV). Four variables were identified as risk factors associated with the introduction of AIV by multivariate logistic regression: 'introduction of end-of-lay chickens ' (odds ratio (OR) = 36.6), 'sharing of farm equipment among farms' (OR = 29.4), 'incomplete hygiene measures of farm visitors on shoes, clothes and hands' (OR = 7.0), and 'direct distance to the nearest case farm' (0-500 m, OR = 8.6; 500-1000 m, OR = 0.8; 1000-1500 m, OR = 20.1; referenced more than 1500 m). We fully believe that strict biosecurity measures should be applied against any incursion points so as not to introduce AIV into more farms.

As-received or shot peened 7075-T7351 single-edged notch bend (SENB) specimens, 8.1-mm thick, were fatigued at a constant maximum load and at stress ratios of R = 0.1 and 0.8 to predetermined numbers of fatigue cycles or to failure. The SENB specimens were then fractured by overload and the tunnelling crack profiles were recorded. The crack-growth rate, da/dN, after crack initiation at the notch was determined by crack-profile measurement and fractography at various fatigue cycles. The shot peened surface topography and roughness was also evaluated by three-dimensional (3-D) laser scanning microscopy. Residual stresses in the as-received specimens and those generated by shot peening at Almen scales of 0.004A, 0.008A, 0.012A and 0.016A, were measured by an X-ray diffraction stress analyser with an X-ray target, CrK, every 0.1 mm to a depth of 1 mm. The 3-D stress intensity factor of the curved crack front was determined by the superposition of the 3-D finite element solutions of the stress intensity factor of the loaded SENB specimen without the residual stress and the stress intensity factor of the unloaded SENB specimen with a prescribed residual stress distribution. da/dN versus the resultant stress intensity factor amplitude, ΔK I , plots showed that while the residual stress locally retarded the crack-growth rate it had no effect on the overall crack-propagation rate.

Background: A definite threshold of the peripheral blood eosinophile count that indicates the presence of hypereosinophilia-associated diseases has not yet been determined. Methods: The threshold eosinophile count at which cases of hypereosinophilia-associated diseases (n = 25) can be differentiated from those of bronchial asthma (n = 101) was determined. Then, the incidences of eosinophile counts greater than 1.0 × 109/l or the threshold level, were studied by analysis of 43 805 samples sent to the laboratory, and the diseases associated with the increased counts were determined. Results: The eosinophile count in cases of hypereosinophilia-associated diseases and in those of bronchial asthma were 10.967 ± 1.680 × 109/l and 0.574 ± 0.045 × 109/l, respectively (P

A new concept for the design and laccase-catalyzed preparation of "artificial urushi" from new urushiol analogues is described. The curing proceeded under mild reaction conditions to produce the very hard cross-linked film (artificial urushi) with a high gloss surface. A new cross-linkable polyphenol was synthesized by oxidative polymerization of cardanol, a phenol derivative from cashew-nut-shell liquid, by enzyme-related catalysts. The polyphenol was readily cured to produce the film (also artificial urushi) showing excellent dynamic viscoelasticity.

Multiple cancer associated with esophageal cancer is not uncommon; however, synchronous esophageal and renal cell carcinoma is very rare. Only three cases have been reported to date, and one of these patients was treated in our institution. We have since successfully treated another patient. Here, we report the two cases treated in our institution. In the first case, esophagectomy, nephrectomy, and reconstruction using a gastric tube were carried out in one stage. Post-operative renal function was temporarily impaired by the complications of anastomotic leakage and pyothorax but no hemodialysis was needed. In the second case, as the patient had undergone distal gastrectomy because of gastric cancer, we chose a two-stage operation, i.e. esophagectomy and nephrectomy as the first stage, followed by reconstruction using a colon substitute after 4 weeks, resulting in only slight renal dysfunction. Patients 1 and 2 are alive and well 7 years and 2 years after the operations respectively.

Switching characteristics such as, rise and decay times and switchabllity with FET, and dynamic luminance of SS (surface stabilized) and PS(polymer stabilized)-FLCDs have been investigated by computer simulation and experiments. The influence of polar anchoring strength on the enhancement of decay process (degradation of memory effect) in SS-FLCD is shown. Rise and decay times of PS-FLCD are shown to be 120 μs, respectively. Dynamic luminance in the impulsive responses of these FLCD technologies are discussed.

The composition of high-density lipoprotein (HDL) changes during inflammation; however, potential changes of HDL function during inflammation and the effects of acute phase proteins that are either on the HDL particles or in the serum have not been clarified. The concentrations of C-reactive protein (CRP), serum amyloid A protein (apoSAA) isoforms, lipids and apolipoproteins, and the activities of lecithin-cholesterol acyltransferase (LCAT) and paraoxonase (PON) were measured before and after laparoscopic cholecystectomy, in 12 patients with cholecystolithiasis to clarify the function of acute-phase HDL and the relationship between acute-phase proteins and HDL functions. Both acute-phase apoSAA (A-apoSAA) and CRP increased, reached their maximum levels 3-6 days after the operation, and then returned to preoperative levels after 2 weeks. In contrast, apolipoproteins and LCAT decreased reciprocally, reached their minimum levels 3-6 days after the operation, and returned to preoperative levels after 2 weeks. However, PON decreased 3-6 days after the operation, and remained low even after 2 weeks. At the nadir the mean activities of LCAT and PON were 56 and 76% of the preoperative levels, respectively. HDL-cholesterol or constitutive apoSAA did not change significantly. LCAT has been reported to be involved in reverse-cholesterol transport and PON to be preventive for lipid peroxidation of low-density lipoprotein in vitro. Thus, during the acute phase of inflammation, HDL may be altered to an atherogenic state due to a decrease in LCAT and PON activities. Therefore, this longitudinal analysis was carried out to determine whether HDL function is modified in a single episode of inflammation and thus may contribute to the occurrence of atherosclerotic disease in patients with chronic or recurrent acute inflammation.

The human optic nerve contains a heterogeneous population of astrocytes. In situ, a specialized subpopulation of astrocytes was distinguished in the adult optic nerve head by expression of neural cell adhesion molecule (NCAM). To further study the biology of astrocytes, we have developed and characterized cells grown from explanted optic nerve heads and myelinated optic nerves as in vitro model systems. Second or third passage cells were processed for immunocytochemistry using antibodies against glial fibrillary acidic protein (GFAP) and cell surface epitopes: CD56/NCAM, HNK-1/NCAM, A2B5, and O4. Synthesis and gene expression of NCAM were characterized by Western blot analysis and RNase protection assay. Cells grown from myelinated optic nerves expressing GFAP, but not NCAM or A2B5, were identified as type 1A astrocytes, and cells expressing GFAP and A2B5, but not NCAM, were identified as type 2 astrocytes. Cells grown from explanted optic nerve head expressing GFAP, NCAM, and O4 were identified as type 1B astrocytes. Expression of NCAM by type 1B astrocytes may provide these cells with adhesion properties that allow them specialized responses in their microenvironment. Astrocytes from the lamina cribrosa may form a functional barrier to prevent myelination of the retina. In glaucoma, these astrocytes may be exposed to stresses due to fluctuation in intraocular pressure and therefore participate in the optic nerve changes associated with glaucomatous optic neuropathy.

Background and hypothesis: Mechanisms of heart failure in elderly hypertensive patients with hypertrophy have not been studied sufficiently. We hypothesized that impaired increment of coronary blood flow in response to increases in heart rate could be responsible for the occurrence or aggravation of heart failure. Methods: To test this hypothesis, we measured coronary hemodynamics and lactate balance during basal conditions and atrial pacing in 21 elderly patients aged ≥ 65 years (mean 74 ± 6 years) without coronary arterial disease: 7 normoten-sive control patients (Group 1), 7 hypertensive hypertrophic patients without a history of congestive heart failure (Group 2), and 7 patients with such history (Group 3). Coronary sinus blood flow (CSBF) was measured in coronary sinus using a thermodilution catheter. Results: During basal conditions, heart rate did not differ among the three groups (67 ± 3 in Group 1,65 ±11 in Group 2, and 63 ± 6 beats/min in Group 3). CSBF was significantly higher in the two hypertrophic groups than in the control group, but CSBF normalized by left ventricular mass was significantly lower in both hypertrophic groups. External mechanical efficiency (EME) obtained as left ventricular work divided by myocardial oxygen consumption did not differ among groups during basal conditions (36 ± 9% in Group 1, 35 ± 8% in Group 2, and 29 ± 9% in Group 3, NS). During atrial pacing to increase heart rate by 25 ± 5% (lower) and 54 ± 6% (higher), the increases in CSBF were markedly limited in both hypertrophic groups, and the response in Group 3 was more depressed than that in Group 2. EME did not change in the control group or in Group 2, but did decrease to 21 ± 5% in Group 3 during the higher pacing rate (p

We have isolated and characterized the Δ9-desaturase gene (Ole1), which codes for a key enzyme involved in regulating membrane fluidity in animal cells and microorganisms, from two strains ofHistoplasma capsulatum, one that is temperature-tolerant (G217B) and the other temperature-susceptible (downs). These pathogenic fungi are dimorphic in that they undergo a morphologic transition from the mycelial to yeast-like form when the temperature of incubation is switched from 25 to 37°C or when they infect a susceptible host. The coding sequences of the two genes, both containing an intron of 93 nucleotides, arevirtually identical and analogous to the Δ9-desaturase gene ofSaccharomyces cerevisiae and those of the rat, mouse and human.Ole1 transcription of the thermotolerant G217B and thermosensitive Downs strains is similar in yeast phase cells and during the temperature shift down from 34, 37, or 40 to 25°C (yeast-to-mycelia transition). Nevertheless, Δ9-desaturase gene is transcriptionally inactive in mycelia of G217B at 25°C while it is actively transcribed in the Downs strain at the same temperature. These results are in agreement with the finding that membranes of the Downs strain have a higher level of oleic acid. The differential expression of Δ9-desaturase genes is discussed in relationship to differences in thermosensitivity in the fungal isolates and in regulating the level of expression of heat shock genes.

A total of 178 sera, including 68 from proven cases of histoplasmosis (65 positive for the presence of Histoplasma capsulatum var. capsulatum antibodies and three positive for antigen), 93 from patients with suspected histoplasmosis but with no laboratory evidence of H. capsulatum var. capsulatum infection, 14 from humans with heterologous fungal and non-fungal infections and three from normal individuals, were tested for IgG H. capsulatum antibodies and M or M and H precipitins by enzyme immunoassay (EIA) (Meridian Diagnostics, Cincinnati, OH, USA) and microimmunodiffusion (MID) respectively. Sixty-three of the 68 histoplasmosis case sera demonstrated IgG antibody, and 65 of 68 demonstrated the presence of specific precipitins in the MID test. Nine positive case sera, when tested with the Laboratory Branch complement fixation (LBCF) test, reacted positively to whole yeast and histoplasmin antigens (titres 1:8 to 1:512). Three histoplasmosis case sera repeatedly tested negative for IgG, specific precipitins and complement-fixing antibodies, whereas they were positive for Histoplasma antigen. Eighteen of 95 sera from patients without evidence of histoplasmosis demonstrated IgG antibody in the EIA only. Among these positive sera, three out of three cases of aspergillosis and three out of five cases of blastomycosis were confirmed. Sera from HIV-infected and healthy individuals did not show IgG or M and/or H antibodies to H. capsulatum. Ninety-three sera were negative by both EIA and MID. The EIA for IgG was less sensitive (97%) than MID (100%). The specificity of EIA and MID was 84% and 100% respectively.

Responses of plasma vasoactive intestinal polypeptide (VIP) to methacholine inhalation and to exercise loading were studied in asthmatic patients to clarity a significant role of the peptidc. The mean of basal VIP in asthmatics was not significantly different from the normals. The levels were increased after FEV (1. 0) (forced expiratory volume in a second) decreased to 80% of the baseline following methacholine inhalation and were returned to values similar to the baseline when FEV (1. 0) recovered to 100%. Thirty minutes after the exercise, mean VIP was significantly lower in patients with the lowest FEV (1. 0). These results suggest that VIPergic system contributes, at least partly, to relax constricted bronchioles in some patients with bronchial asthma.

The Friedel-Crafts acylation of substituted benzenes proceeds smoothly by using a catalytic amount of a lanthanide trifluoromethanesulfonate [Ln(OTf)3], which is easily recovered from the reaction mixture and reused without a decrease in catalytic activity.

The aldol reaction of silyl enolates with aldehydes or acetals using a lanthanide trifluoromethanesulfonate as catalyst smoothly proceed in organic solvents to afford the corresponding aldol-type adducts in high yields. The catalyst can be easily recovered after the reaction is completed, and yields of the 2nd run of the catalyst are comparable to those of the 1st run

The Michael reaction of silyl enolates with α,β-unsaturated ketones and the Diels-Alder reaction of carbonyl containing dienophiles are catalyzed by a catalytic amount of lanthanide trifluoromethansulfonate to give the corresponding adducts in high yields. In both reactions, the catalyst can be recovered and reused.

Novel silyl enolates, prepared in situ from ketones and dimethylsilyl ditriflate (Me 2 Si(OTf) 2 ) in the presence of a tertiary amine, reacted smoothly with electrophiles such as aldehydes, acetals, or α,β-unsaturated ketones without catalyst at -78 o C to afford the corresponding adduct in high yields and with high selectivities

Allyltrichlorosilanes regioselectively react with aldehydes in N,N-dimethylformamide (DMF) without catalyst to afford the corresponding homoallylic alcohols in high yields. Syn and anti homoallylic alcohols are stereoselectively obtained from (Z)- and (E)-crotyltrichlorosilane, respectively, under neutral conditions.

In the presence of a catalytic amount of chiral ytterbium trifluoromethanesulfonate (triflate), acyl-1,3-oxazolidin-2-ones or methacrolein reacted with cyclopentadiene to afford the corresponding Diels-Alder adduct in high yields and with moderate to excellent enantiomeric excesses.

Highly diastereo- and enantioselective aldol-type reactions of a silyl enol ether with aldehydes including aromatic, aliphatic, and α,β-unsaturated ones are performed in propionitrile (solvent) by the use of a catalytic amount of chiral tin(II) Lewis acid consisting of tin(II) triflate and a chiral diamine. The asymmetric environments created by the chiral catalysts in the enantioselective aldol reaction are discussed by employing five kinds of chiral diamines.

6-Deoxy-D-allose is prepared from simple achiral starting materials, butynal and chloroacetic acid, via two key steps of the chiral tin(II) triflate mediated asymmetric aldol reaction and the dihydroxylation using osmium tetroxide.