Strong photo-reductants have applications in photo-redox organic synthesis involving reductive activation of C-X(halide) and C=O bonds. We report herein air-stable PtII complexes supported by tetradentate bis(phenolate-NHC) ligands having peripheral electron-donating N-carbazolyl groups. Photo-physical, electrochemical, and computational studies reveal that the presence of N-carbazolyl groups enhances the light absorption and redox reversibility because of its involvement into the frontier MOs in both ground and excited states, making the complexes robust strong photo-reductant with E([Pt]+/ *) over -2.6 V vs. Cp2 Fe+/0 . The one-electron reduced [Pt]- species are stronger reductants with EPC ([Pt]0/- ) up to -3.1 V vs. Cp2 Fe+/0 . By virtue of the strong reducing nature of these species generated upon light excitation, they can be used in light-driven reductive coupling of carbonyl compounds and reductive debromination of a wide range of unactivated aryl bromides.

Microtubules are cytoskeletal structures critical for mitosis, cell motility, and protein and organelle transport, and are a validated target for anticancer drugs. However, how tubulins are regulated and recruited to support these distinct cellular processes is incompletely understood. Post-translational modifications of tubulins are proposed to regulate microtubule functions and dynamics. Although many of these modifications have been investigated, only one prior study reports on tubulin methylation and an enzyme responsible for this methylation. Here, we used in vitro radiolabelling, mass spectrometry, and immunoblotting approaches to monitor protein methylation, and immunoprecipitation, immunofluorescence, and pull-down approaches to measure protein-protein interactions. We demonstrate that N-lysine methyltransferase 5A (KMT5A or SET8/PR Set7), which methylates lysine 20 in histone H4 (H4K20), bound α-tubulin and methylated it at a specific lysine residue, Lys-311. Furthermore, LSF/CP2, a known transcription factor, bound both α-tubulin and SET8, and enhanced SET8-mediated α-tubulin methylation in vitro In addition, we found that the ability of LSF to facilitate this methylation is countered by factor quinolinone inhibitor 1 (FQI1), a specific small-molecule inhibitor of LSF. These findings suggest the general model that microtubule-associated proteins, including transcription factors, recruit or stimulate protein-modifying enzymes to target tubulins. Moreover, our results point to dual functions for both SET8 and LSF, not only in chromatin regulation, but also in cytoskeletal modification.

Aberrant activation of beta-catenin/TCF (T-cell factor) signaling is frequently observed in the pancreatic cancer. However, the regulation of nuclear beta-catenin/TCF transcription machinery remains largely unknown. In this study, TFCP2 (transcriptional factor CP2) expression in pancreatic cancer was detected by qPCR, immunohistochemistry and western blot. Western blot, colony formation assay, migration and invasion experiment were performed to investigate the effects of TFCP2 on the growth and migration of pancreatic cancer cells. In vivo, mouse metastasis models were utilized to determine metastasis ability. Western blots were used to evaluate the related protein expression. Luciferase reporter assay was used to explore the role of TFCP2 on beta-catenin/TCF signaling. We have shown that the transcription factor TFCP2 was up-regulated in the pancreatic cancer. Over-expression of TFCP2 promoted the growth, migration, invasion and colony formation of pancreatic cancer cells, while knocking down the expression of TFCP2 inhibited the growth, migration, invasion, colony formation and metastasis of pancreatic cancer cells. The mechanism study revealed that TFCP2 interacted beta-catenin, enhanced the interaction between beta-catenin and TCF4, and activated beta-catenin/TCF signaling. Taken together, our study demonstrated the oncogenic roles of TFCP2 in pancreatic cancer, and suggested that TFCP2 might be a target for the treatment of pancreatic cancer.

We analysed hyaline cartilage of human knee and ankle joints for collagen and proteoglycan turnover in order to find differences in the metabolism and biochemical content of the extracellular matrix that could explain the higher prevalence of osteoarthritis (OA) in the knee joint, compared to the ankle joint.

Cartilage tissue from ankle and knee joints of OA patients were assessed for total collagen and proteoglycan content. For turnover, the aggrecan 846-epitope (CS 846), the type II collagen C-propeptide (CP2) and the collagenase-generated intrahelical cleavage neoepitope (C2C) were quantified.

Molecular analyses showed that type II collagen turnover (CP2 and C2C) was significantly elevated in the ankle, whereas aggrecan turnover (CS 846), total proteoglycan and total collagen were comparable between both joints. Analysis of the inter-relationships in the components of cartilage matrix turnover showed a significant positive correlation of C2C vs CP2.

The data suggest an increased type II collagen turnover in ankle vs knee OA cartilage but a comparable aggrecan turnover and comparable contents of type II collagen and proteoglycan. These findings point towards a focused attempt in advanced OA cartilage to structurally repair the collagen network that was more pronounced in the ankle joint and may explain in part the higher prevalence of OA in the knee as compared to the ankle joint.

Complement-mediated killing of pathogens through lytic pathway is an important effector mechanism of innate immune response. C9 is the ninth member of complement components, creating the membrane attack complex (MAC). In the present study, a putative cDNA sequence encoding the 650 amino acids of C9 and its genomic organization were identified in grass carp Ctenopharyngodon idella. The deduced amino acid sequence of grass carp C9 (gcC9) showed 48% and 38.5% identity to Japanese flounder and human C9, respectively. Domain search revealed that gcC9 contains a LDL receptor domain, an EGF precursor domain, a MACPF domain and two TSP domain located in the N-terminal and C-terminal, respectively. Phylogenetic analysis demonstrated that gcC9 is clustered in a same clade with Japanese flounder, pufferfish and rainbow trout C9. The gcC9 gene consists of 11 exons with 10 introns, spacing over approximately 7 kb of genomic sequence. Analysis of gcC9 promoter region revealed the presence of a TATA box and some putative transcription factor such as C/EBP, HSF, NF-AT, CHOP-C, HNF-3B, GATA-2, IK-2, EVI-1, AP-1, CP2 and OCT-1 binding sites. The first intron region contains C/EBPb, HFH-1 and Oct-1 binding sites. RT-PCR and Western blotting analysis demonstrated that the mRNA and protein of gcC9 gene have similar expression patterns, being constitutively expressed in all organs examined of healthy fish, with the highest level in hepatopancreas. By real-time quantitative RT-PCR analysis, gcC9 transcripts were significantly up-regulated in head kidney, spleen, hepatopancreas and down-regulated in intestine from inactivated fish bacterial pathogen Flavobacterium columnare-stimulated fish, demonstrating the role of C9 in immune response.

BACKGROUND

We have shown that a low glutathione concentration and synthesis rate in erythrocytes are associated with a shortage of protein-derived cysteine in children with edematous severe acute malnutrition (SAM).

OBJECTIVE

We tested the hypothesis that methionine supplementation may increase protein-derived cysteine and upregulate cysteine synthesis, thereby improving glutathione synthesis during the early treatment of edematous SAM.

METHODS

The cysteine flux, its de novo synthesis and release from protein breakdown, and erythrocyte glutathione synthesis rate were measured in 12 children with edematous SAM in the fed state by using stable isotope tracers at 3 clinical phases as follows: 3 ± 1 d (±SE) [clinical phase 1 (CP1)], 8 ± 1 d [clinical phase 2 (CP2)], and 14 ± 2 d (clinical phase 3) after admission. Subjects were randomly assigned to receive equimolar supplements (0.5 mmol ⋅ kg(-1) ⋅ d(-1)) of methionine or alanine (control) immediately after CP1.

RESULTS

In the methionine compared with the alanine group, cysteine flux derived from protein breakdown was faster at CP2 than CP1 (P < 0.05), and the change in plasma cysteine concentration from CP1 to CP2 was greater (P < 0.05). However, there was no evidence of a difference in cysteine de novo synthesis and its total flux or erythrocyte glutathione synthesis rate and concentration between groups.

CONCLUSIONS

Methionine supplementation increased cysteine flux from body protein but had no significant effect on glutathione synthesis rates. Although cysteine is made from methionine, increased dietary cysteine may be necessary to partially fulfill its demand in edematous SAM because glutathione synthesis rates and concentrations were less than previous values shown at full recovery. This study was registered at clinicaltrials.gov as NCT00473031.

Lower-limb exoskeletons have been used in gait rehabilitation to facilitate the restoration of motor skills. These robotics systems could be complemented by Brain-Computer Interfaces (BCIs) to assist or rehabilitate people with walking disabilities. In this preliminary study, electroencephalography-based brain functional connectivity is analyzed during exoskeleton-assisted gait motor imagery (MI) training. Partial Directed Coherence (PDC) analysis was employed to assess the exchange of information flow between EEG signals during gait MI in four healthy subjects, two using an exoskeleton and two without using it. Besides, in order to explore the functional connectivity, an outflow index based on the number of significant directed connectivities revealed by the PDC analysis is proposed. We found that the outflow index increases in the central zone (C2, C3, C4) while decreases in the central-parietal (CP1, CP2) and fronto-central (FC1) zones when the training was assisted by an exoskeleton. The results obtained can be useful to obtain informative features for BCI applications as well as in motor rehabilitation.

The present study aimed to investigate the effect of stage duration in incremental protocols on lactate threshold (LT), determined by different methods. Sixteen semi-professional soccer-players performed a 4-min stage incremental discontinuous (DP) and two maximal incremental running continuous (1 km h-1· min-1, CP1; and 1 km h-1·2 min-1 CP2) protocols. Blood-lactate concentration [La-] was measured at baseline and during the protocols. LT was determined using DMAX, DMAX-MOD, 4-mM⋅L-1, Δ1-mM⋅L-1 and Log-Log methods. Log-Log showed no difference in LT between CP1, CP2 and DP. Conversely, LT was determined at higher velocity in CP1 than CP2 for DMAX (15.2 ± 0.5 vs 14.4 ± 1.2 km⋅h-1, P = 0.002), DMAX-MOD (16.0 ± 0.5 vs 14.7 ± 1.3 km⋅h-1, P < 0.001), 4-mM⋅L-1 (15.5 ± 1.4 vs 14.4 ± 1.2 km⋅h-1, P < 0.001), Δ1-mM⋅L-1 (15.5 ± 1.3 vs 14.4 ± 1.2 km⋅h-1, P < 0.001). Higher LT in CP1 than DP for DMAX (15.2 ± 0.5 vs 13.0 ± 1.0 km⋅h-1, P < 0.001) and DMAX-MOD (16.0 ± 0.5 vs 13.6 ± 1.6 km⋅h-1, P < 0.001) was found (P < 0.001). Log-Log resulted in shorter but accurate protocols to determine LT.

This study evaluated the performance, gut microbiota, and blood metabolites in broiler chickens fed cranberry and blueberry products for 30 days. A total of 2,800 male day-old broiler Cobb-500 chicks were randomly distributed between 10 diets: control basal diet; basal diet with bacitracin (BACI); four basal diets with 1 and 2% of cranberry (CP1, CP2) and blueberry (BP1, BP2) pomaces; and four basal diets supplemented with ethanolic extracts of cranberry (COH150, COH300) or blueberry (BOH150, BOH300) pomaces. All groups were composed of seven replicates (40 birds per replicate). Cecal and cloacal samples were collected for bacterial counts and 16S rRNA gene sequencing. Blood samples and spleens were analyzed for blood metabolites and gene expressions, respectively. The supplementation of COH300 and BOH300 significantly increased the body weight (BW) during the starting and growing phases, respectively, while COH150 improved (P < 0.05) the overall cumulated feed efficiency (FE) compared to control. The lowest prevalence (P = 0.01) of necrotic enteritis was observed with CP1 and BP1 compared to BACI and control. Cranberry pomace significantly increased the quinic acid level in blood plasma compared to other treatments. At days 21 and 28 of age, the lowest (P < 0.05) levels of triglyceride and alanine aminotransferase were observed in cranberry pomace and blueberry product-fed birds, respectively suggesting that berry feeding influenced the lipid metabolism and serum enzyme levels. The highest relative abundance of Lactobacillaceae was found in ceca of birds fed CP2 (P < 0.05). In the cloaca, BOH300 significantly (P < 0.005) increased the abundances of Acidobacteria and Lactobacillaceae. Actinobacteria showed a significant (P < 0.05) negative correlation with feed intake (FI) and FE in COH300-treated birds, whereas Proteobacteria positively correlated with the BW but negatively correlated with FI and FE, during the growing phase. In the spleen, cranberry products did not induce the release of any pro-inflammatory cytokines but upregulated the expression of several genes (IL4, IL5, CSF2, and HMBS) involved in adaptive immune responses in broilers. This study demonstrated that feed supplementation with berry products could promote the intestinal health by modulating the dynamics of the gut microbiota while influencing the metabolism in broilers.

Keywords: blood metabolites; broilers; cranberry and blueberry pomaces; gut microbiota; immunity; spleen.

Viruses are a major biotic constraint on sweet potato (Ipomoea batatas (L.) Lam) production worldwide. In 2005, 10 to 60% viral disease incidence was observed in sweet potato fields. Symptoms include ring and chlorotic spots, puckering, feathering, vein clearing, and leaf curl with chlorotic specks and pink spots. Cuttings from symptomatic plants were collected from Kerala (two clones), Orrisa (eight clones), and Adrapradesh (three clones) and maintained in an insect-proof glasshouse. Leaves from symptomatic plants were mechanically inoculated to I setosa, I. nil, Nicotiana tabacum, N. benthamiana, Datura stramonium, and Chenapodium quinoa (12 seedlings each). Vein clearing, netting, and leaf distortion were observed in I. setosa and N tabacum 7 days postinoculation, chlorotic spots observed in N. benthamiana, and violet spots and violet margins on leaves observed on I. Nil. No symptoms were observed on D. stramonium and C. quinoa. When scions from the symptomatic sweet potato plants were graft inoculated onto I. setosa, vein clearing, leaf curl, and puckering-like symptoms were observed within 5 days. Mosaic and leaf curling symptoms were also observed on mechanically inoculated N. tabacum. Total nucleic acids isolated from the 33 field-collected sweet potato samples, graft inoculated I. setosa plants, and mechanically inoculated N. tabacum and I. nil plants were used for PCR and reverse transcription (RT)-PCR with geminivirus group specific (2) and potyvirus group specific primers (1). The expected 530-bp and 1.3-kb fragment were generated from the geminivirus and potyvirus primer sets, respectively. Potyvirus alone was detected in 7 of the 33 field-collected plants; geminivirus alone was detected in 7 other plants, while 19 plants contained detectible levels of potyvirus and geminivirus. To further identify the viruses, nested primers specific for the coat protein gene of Sweet potato feathery mottle virus (SPFMV) (CP1S 5'AGT GGG AAG GCA CCA TAC ATA GC 3', CP1A5' GCA GAG GAT GTC CTA TTG CAC ACC 3') (CP2S 5'TCT AGT GAA CGT ACT GAA TTC AAA GA 3', CP2A 5'ATT GCA CAC CCC TGA TTC CTA AGA 3') and Sweet potato leaf curl virus (SPLCV) (CP1- 5'ATG ACA GGG CGA ATT CGC GTT TC 3', CP2- 5'TTA ATT TTT GTG CGA ATC ATA 3') were designed. I. setosa and N. tabacum were amplified with SPFMV and SPLCV primers and the amplicons of 960 and 764 bp, respectively, obtained were subsequently cloned into pGEM-T Easy vector and sequenced. Nucleotide BLAST analysis revealed that the 960-bp fragment (GenBank Accession No. EF015398.) was 98% identical to two Egyptian isolates of SPFMV (Nos. AJ 515379 and AJ 515378). The nucleotide sequence of the 764-bp products (Nos. EF 151926 and EF15483) from the samples collected from Kerala and Orisa was 95% identical to each other. The sequence identity of EF 15483 with Sweet potato leaf curl Georgia virus (SPLCGV) isolate AF326775. was 91% and identity with China isolate DQ 512731 was 90% The isolate EF 151926 also was 91% identical to the SPLCGV with a high query and alignment score whereas identity with the China isolate was 91% with a low query coverage and alignment score. Phylogenic analysis with MEGA software program also showed the highest sequence similarity with SPLCGV, hence it is concluded that the geminivirus isolate under study is SPLCGV. To our knowledge, this is the first report of identification of SPFMV and SPLCGV occurring on sweet potato in India. Further study is required to understand the consequences of occurrence of these two viruses in India. References: (1) D. Colinet et al. Plant Dis. 28:223 1998. (2) D. D. Deng et al. Ann. Appl. Biol 125:327, 1993.

TFCP2L1 (transcription factor CP2-like 1) is a transcriptional regulator critical for maintaining mouse and human embryonic stem cell (ESC) pluripotency. However, the direct TFCP2L1 target genes are uncharacterized. Here, using gene overexpression, immunoblotting, quantitative real-time PCR, ChIP, and reporter gene assays, we show that TFCP2L1 primarily induces estrogen-related receptor β (Esrrb) expression that supports mouse ESC identity and also selectively enhances Kruppel-like factor 4 (Klf4) expression and thereby promotes human ESC self-renewal. Specifically, we found that in mouse ESCs, TFCP2L1 binds directly to the Esrrb gene promoter and regulates its transcription. Esrrb knockdown impaired Tfcp2l1's ability to induce interleukin 6 family cytokine (leukemia inhibitory factor)-independent ESC self-renewal and to reprogram epiblast stem cells to naïve pluripotency. Conversely, Esrrb overexpression blocked differentiation induced by Tfcp2l1 down-regulation. Moreover, we identified Klf4 as a direct TFCP2L1 target in human ESCs, bypassing the requirement for activin A and basic fibroblast growth factor in short-term human ESC self-renewal. Enforced Klf4 expression recapitulated the self-renewal-promoting effect of Tfcp2l1, whereas Klf4 knockdown eliminated these effects and caused loss of colony-forming capability. These findings indicate that TFCP2L1 functions differently in naïve and primed pluripotency, insights that may help elucidate the different states of pluripotency.

BACKGROUND

Allogeneic hematopoietic cell transplantation (HCT) was a standard therapy in chronic phase (CP) chronic myeloid leukemia (CML). As a result of the effective therapy with tyrosine kinase inhibitors (TKI), HCT was shifted to defined clinical situations. We present the results of observational prospective analysis of 28 CML patients undergoing HCT after exposure to, at least, two lines of TKI (including dasatinib and/or nilotinib), with respect to response, overall survival (OS), treatment toxicity, graft versus host disease (GVHD), and progression/relapse incidence.

RESULTS

All the patients but one engrafted with median time 19 days. OS for patients in CP1 and CP2/accelerated phase (AcP) were 92.9 and 85.7 %, respectively. Six patients allotransplanted in blast crisis (BC) CML died early after HCT. Eighteen patients achieved deep molecular remission (MR(4.5) or MR(4.0)). Relapse incidence was 29.6 %. Median time to progression (TTP) differs significantly depending on the CML phase prior to HCT, the best response achieved after HCT and development of chronic GvHD. NRM yielded the values 7.1, 12.5, and 50 % in CP1, CP2/AcP, and BC, respectively. Fatal outcome, due to veno-occlusive disease (VOD), was observed in two (7 %) patients. In five (17.9 %) patients, mild or moderate VOD was observed with no negative impact of preceding therapy with TKI2. Acute GvHD was diagnosed in 25.9 % of patients, while chronic GvHD developed in 42.9 % of individuals.

CONCLUSIONS

Pretransplantation therapy with TKI2 in CP CML is safe and reasonable. In BC, the optimal approach before HCT is to reduce the leukemic burden and achieve subsequent CP.

In the present study, the emphasis is laid on the self aggregation behavior of copper based inorganic-organic hybrids in aqueous media. The two complexes, cationic hexadecyl pyridinium trichloro cuprate (1 : 1), [Cp](+)[CuCl3](-), and bishexadecylpyridinium tetrachloro cuprate (2 : 1), [Cp2](2+)[CuCl4](2-), were synthesized using the ligand insertion method. The complexes were characterized using elemental analysis, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), nuclear magnetic resonance (NMR) and thermogravimetric analysis. The copper complexes were found to be thermally stable, and in the solid state, they possessed the perovskite arrangement with [Cp2](2+)[CuCl4](2-) exhibiting superior stability and crystallinity. The self aggregation behavior of the prepared complexes was analyzed in solution phase (in aqueous medium) using surface tension, conductivity, XRD and small angle neutron scattering (SANS). The results show that the presence of copper as a co-ion in both the stoichiometries results in lower critical micellization concentrations than their precursor. Micellization was thermodynamically spontaneous and micelles formed were ellipsoidal in shape and underwent a prolate ellipsoidal growth with an increase in the concentration of metallosurfactant, as estimated from the SANS. Furthermore, these metallosurfactants were investigated for biocompatibility (using hemolytic assay), antimicrobial activity (fungus and bacteria) and cytotoxicity using human cancerous cells. The hemolysis activity was found to depend on the aggregated state of the metallosurfactants, displaying the highest activity in the monomeric state, and the minimum for post micellar concentrations. The surfactants were found to enhance the antibacterial activity by twofold or more, with the addition of metal in both the stoichiometries. On the contrary, for anticancer and antifungal activities, barely any regular trend or generalization could be obtained. Nevertheless, the copper complexes exhibited high IC50 values for fR2 (healthy cells) signifying their higher safety in comparison to the cancerous cells.

When individual neurons in a circuit contain multiple neuropeptides, these peptides can target different sets of follower neurons. This endows the circuit with a certain degree of flexibility. Here we identified a novel family of peptides, the Aplysia SPTR-Gene Family-Derived peptides (apSPTR-GF-DPs). We demonstrated apSPTR-GF-DPs, particularly apSPTR-GF-DP2, are expressed in the Aplysia CNS using immunohistochemistry and MALDI-TOF MS. Furthermore, apSPTR-GF-DP2 is present in single projection neurons, e.g., in the cerebral-buccal interneuron-12 (CBI-12). Previous studies have demonstrated that CBI-12 contains two other peptides, FCAP/CP2. In addition, CBI-12 and CP2 promote shortening of the protraction phase of motor programs. Here, we demonstrate that FCAP shortens protraction. Moreover, we show that apSPTR-GF-DP2 also shortens protraction. Surprisingly, apSPTR-GF-DP2 does not increase the excitability of retraction interneuron B64. B64 terminates protraction and is modulated by FCAP/CP2 and CBI-12. Instead, we show that apSPTR-GF-DP2 and CBI-12 increase B20 excitability and B20 activity can shorten protraction. Taken together, these data indicate that different CBI-12 peptides target different sets of pattern-generating interneurons to exert similar modulatory actions. These findings provide the first definitive evidence for SPTR-GF's role in modulation of feeding, and a form of molecular degeneracy by multiple peptide cotransmitters in single identified neurons.

The present study describes transcription of mRNA from genes encoding metabolic or structural proteins during excystation of Cryptosporidium parvum oocysts. RNA was harvested from C. parvum oocysts before excystation, and at 5, 10, and 15 min during excystation. Subtractive cDNA libraries were prepared by using mRNA from non-excysted C. parvum oocysts to "subtract out" mRNA from excysting oocysts. The "subtracted" cDNA was used to prepare libraries enriched for transcripts possibly involved in excystation. From these libraries, over 1,000 expressed sequence tags (ESTs) were analyzed by DNA sequencing followed by BLAST-N and BLAST-X analysis. While several gene products involved in cell metabolism and cell signaling were consistently recovered, transcription levels, as reflected by the relative number of cDNA sequences (19.2% total), were highly up-regulated in genes coding for structural proteins such as Cp2, CpTSP, CpHC10, and CpSAg. Moreover, of the greater than 1,000 clones analyzed, a high percentage (12.3%) of ESTs detected in excysting oocysts were for hypothetical C. parvum proteins (CpHyP), whose functions are presently unknown.

Seven cuticle chitin-binding proteins (Ld-CP1v1 to 7) were deduced from antenna cDNAs of adult Colorado potato beetles, Leptinotarsa decemlineata (Say), based on their consensus sequences. The mature proteins consisted of 87-188 residues. Ld-CP1v1 formed a distinct orthologous protein cluster (OP1) along with four proteins from other insect species in a neighbor-joining phylogenetic tree. These proteins also contained a proline glutamine-rich (PQ-rich) region and a highly conserved C-terminal motif (Phr). Their consensus region lacked the defined aromatic triad. Ld-CP2 to 6 clustered with those bearing RR-1 consensus and Ld-CP7 with RR-2 consensus. Ld-CP1v1 to 4 were expressed at the post-ecdysial period in all the developmental stages whereas Ld-CP5 to 7 were expressed mainly in adults.

Exercise critical power (CP) has been shown to represent the highest sustainable work rate (WR) in patients with chronic obstructive pulmonary disease (COPD). Parameter estimation, however, depends on 4 high-intensity tests performed, on different days, to the limit of tolerance (T(lim)). In order to establish a milder protocol that would be more suitable for disabled patients, we contrasted CP derived from 4, 3 and 2 tests (CP4, CP3 and CP2) in 8 males with moderate COPD. In addition, CP was calculated from 2 single-day tests performed on an inverse sequence (CP(2AB) and CP(2BA)): CP values within 5 W from CP4 were assumed as "clinically-acceptable" estimates. We found that [CP4-CP3] and [CP4-CP2] differences were within 5 W in 8 and 6 patients, respectively (95% confidence interval of the differences = -1.3 to 3.5 W and -11.5 to 6.5 W). There was a systematic decline on T(lim) when an exercise bout was performed after a previous test on the same day (P<0.05). Consequently, substantial differences were found between CP4 and any of the CP estimates obtained from single-day tests. In conclusion, clinically-acceptable estimates of CP can be obtained by using 3 or, in most circumstances, 2 constant WR tests in patients with moderate COPD--provided that they are not performed on the same day.

The actin cytoskeleton of host cells has been implicated in Cryptosporidium invasion. However, the underlying mechanism of how actin filaments and associated proteins modulate this process remains unclear. In this study, we use in vitro cultured cell lines, human ileocecal adenocarcinoma HCT-8 and Chinese hamster ovary (CHO), and an in vivo mouse model to investigate the roles of tropomyosin isoforms in Cryptosporidium invasion. Using isoform-specific monoclonal antibodies, we found that the major human tropomyosin (hTM) isoforms expressed in HCT-8 cells are hTM4 and hTM5. HCT-8 cells also express hTM1 at low levels but not hTM2 and hTM3. During Cryptosporidium parvum infection, hTM5 colocalized to the infection sites with a novel parasite membrane protein, CP2. Neither hTM1 nor hTM4 accumulated at infection sites. Similarly, a high level of TM5 and varying amounts of TM4 accumulated at the C. parvum infection sites in CHO cells. CHO cells overexpressing hTM5 exhibit a significantly higher percent of mature meronts early in the infection process relative to CHO cells or CHO cells overexpressing a tropomyosin mutant, chimeric isoform hTM5/3. These results suggest that functional TM5 enhances Cryptosporidium invasion of host cells. In C. parvum-infected mice, accumulation and rearrangement of TM5 and TM4 were detected throughout the infected ileum. Similarly, in the Cryptosporidium muris-infected mice, TM5 accumulated in discrete regions of the epithelial cells of gastric glands and in the oocyst-laden stomach gland lumen. Cryptosporidium infection appears to rearrange and recruit host TM isoforms in both culture cells and in the mouse. Localized accumulation of tropomyosin at the infection sites may facilitate parasite invasion.

OBJECTIVE

The contingent negative variation (CNV) is a widespread electroencephalographic (EEG) potential that occurs during the interval between a warning stimulus and a subsequent imperative stimulus if a mental or motor response is required. The present study was designed to explore the impact of the previous trial on the CNV of the forthcoming trial, that is, how a previous movement affects brain activation preparing the next movement. Effects of alteration of finger (from index to middle, and vice versa) and hand (from left to right, and vice versa) were examined independently from each other.

METHODS

CNV was recorded in 20 right-handed healthy subjects with electrodes placed at F7, F5, F3, F4, F6, F8, FC5, FC3, FC1, FC2, FC4, FC6, T7, C5, C3, C1, C2, C4, C6, T8, CP5, CP1, CP2, CP6, P7, P3, P4 and P8. In a visual/visual S1-choice paradigm, an earlier informative (S1) stimulus which instructed for side and finger of the following movement was followed 3 s later by an imperative (S2) stimulus providing the command to move. Subjects had to respond to each imperative stimulus with an appropriate button press made by brisk flexion movements with the index or middle finger of each hand. The CNV recorded in the interval between the informative and the imperative stimulus was analysed with respect to finger and hand of the present and the preceding movement.

CONCLUSIONS

(1) A change of the side of movement is associated with a widespread increase of negativity contralateral to the currently prepared movement. (2) A change of finger is associated with a focal increase of negativity contralateral to the side of the current movement over temporoparietal and mid-parietal areas. (3) A change of finger results in a widespread increase of negativity over the left hemisphere.

A complex interplay between the catalyst concentration and the reaction rate exists, as shown by kinetic studies, for the [Cp2 TiMe2 ]-catalyzed intermolecular hydroamination of alkynes. The reason for this is a reversible dimerization of the catalytically active species [Eq. (1)]. L1 , L2 =Cp (cyclopentadienyl), Tol-NH (Tol=4-MeC6 H4 , tolyl).

OBJECTIVE

To analyze the outcome of allogeneic bone marrow transplantation (allo-BMT) performed for chronic myeloid leukemia (CML) patients.

METHODS

One hundred and eighteen CML patients received allo-BMT with conditioning regimens of TBI or modified BU/CY. The pre-BMT situations were, 91 cases in first chronic phase (CP1), 19 in accelerating phase (AP), 2 in blast crisis (BC) and 6 in advanced CP >> CP2).

RESULTS

One hundred and nine patients were successfully engrafted. The probabilities of 5 year survival for CP1 and AP patients were 69.6% and 51.0%, and 5 year relapse rates were 3.2% and 12.5%, respectively.

CONCLUSIONS

Disease stage, conditioning regimen and splenomegaly did not affect the time for engraftment. For patients in CP1, splenomegaly was positively correlated with relapse. There was no difference between the effect of the two conditioning regimens on the prognosis.

To investigate the potential role of Sm-Ph species as intermediates in the samarium-catalyzed redistribution of PhSiH3 to Ph2SiH2 and SiH4, the samarium phenyl complex [Cp*2SmPh]2 (1) was prepared by oxidation of Cp2*Sm (2) with HgPh2. Compound 1 thermally decomposes to yield benzene and the phenylene-bridged disamarium complex Cp*2Sm(mu-1,4-C6H4)SmCp*2 (3). This decomposition reaction appears to proceed through dissociation of 1 into monomeric Cp*2SmPh species which then react via unimolecular and bimolecular pathways, involving rate-limiting Cp* metalation and direct C-H activation, respectively. The observed rate law for this process is of the form: rate = k1[1] + k2[1]2. Complex 1 efficiently transfers its phenyl group to PhSiH3, with formation of Ph2SiH2 and [Cp*2Sm(mu-H)]2 (4). Quantitative Si-C bond cleavage of C6F5SiH3 is effected by the samarium hydride complex 4, yielding silane and [Cp*2Sm(mu-C6F5)]2 (5). In contrast, Si-H activation takes place upon reaction of 4 with o-MeOC6H4SiH3, affording the samarium silyl species [structure: see text] Cp*2SmSiH2(o-MeOC6H4) (7). Complex 7 rapidly decomposes to [Cp*2Sm(mu-o-MeOC6H4)]2 (6) and other samarium-containing products. Compounds 5 and 6 were prepared independently by oxidation of 2 with Hg(C6F5)2 and Hg(o-MeOC6H4)2, respectively. The mechanism of samarium-mediated redistribution at silicon, and chemoselectivity in sigma-bond metathesis reactions, are discussed.

In the present investigation, the bioefficacy of developed carbofuran formulations, with PEG-600 (7a, CP1) & PEG-900 (7b, CP2) @ 5, 10 and 20 ppm, along with commercial formulation of carbofuran 3G (CP0) were evaluated against the root-knot nematode, Meloidogyne incognita infecting tomato (cv. Pusa Ruby) in pot and field conditions. The bioefficacy data indicated that the formulations developed by utilizing polymers having PEG - 900 (7b) as hydrophilic segment were effective even at 14 days post inoculation (dpi) as evident from shoot and root length. Also, the reduction in penetration was found to be maximum with CP2 (3.6 - 4.6 J2s) at all concentrations compared to CP1 (6.6-16.4 J2s) and CP0 (29.3-32.6 J2s). Overall, CP2 was more effective in reducing the number of nematodes up to 14 days, compared to CP1 and CP0. Both the CR formulations (CP1 and CP2) in general significantly reduced the number of galls, when compared to CP0. However, under field conditions, lower concentrations (5, and 10 ppm) of CP2, were less effective in controlling the gall formation whereas, CP2 at 20 ppm, was most effective than other treatments. The study revealed that the developed CR formulations of carbofuran have the potential for effective management of M. incognita in tomato under field conditions.

Asp-Phe-Leu-Ala-Glu (DE5) is a frequent sequence of many toxic middle molecular peptides that accumulate in uremic patients. To eliminate these peptides by hemoperfusion, three adsorbents (CP1-Zn(2+), CP2-Zn(2+), and CP3-Zn(2+)) were designed on the basis of coordination and hydrophobic interactions. Adsorption experiments indicated that CP2-Zn(2+) had the highest affinity for DE5 among these three adsorbents. Also, the adsorption capacity of CP2-Zn(2+) in DE5 and DE5-containing peptides was about 2-6 times higher than that of peptides without the DE5 sequence. Linear polymers bearing the same functional groups of the adsorbents were used as models to study the adsorption mechanism via isothermal titration calorimetry (ITC) and computer-aided analyses. The results indicated that coordination and hydrophobic interactions played the most important roles in their affinity. When two carboxyl moieties on Asp and Glu residues coordinated to CP2-Zn(2+), the hydrophobic interaction took place by the aggregation of the hydrophobic amino acid residues with phenyl group on CP2-Zn(2+). The optimal collaboration of these interactions led to the tight binding and selective adsorption of DE5-containing peptides onto CP2-Zn(2+). These results may provide new insight into the design of affinity adsorbents for peptides containing DE5-like sequences.