RESUMEN

Increasing continuity in Dutch maternity care is considered pivotal to improve safety and client-centeredness. Closer collaboration between the historically relatively autonomous professionals and organizations in maternity care is deemed conditional to reach this goal, both by maternity care professionals and policy makers. Governmental policy therefore strives for organizational and financial integration. One of the policy measures has been to stimulate interprofessional and interorganizational collaboration through local obstetric partnerships. This study aimed to gain insight into whether this policy measure supported professionals in reaching the policy aim of increasing integration in the maternity care system. We therefore conducted 73 semistructured interviews with maternity care professionals in the region Northwest Netherlands, from 2014 to 2016. Respondents expressed much willingness to intensify interprofessional and interorganizational collaboration and experienced obstetric partnerships as contributing to this. As such, stimulating integration through obstetric partnerships can be considered a suitable policy measure. However, collaborating within the partnerships simultaneously highlighted deep-rooted dividing structures (organizational, educational, legal, financial) in the maternity care system, especially at the systemic level. These were experienced to hinder collaboration, but difficult for the professionals to influence, as they lacked knowledge, skills, resources and mandate. A lack of clear and timely guidance and support from policy, counterbalancing these barriers, limited partnerships' potential to unify professionals and integrate their services.

Asunto(s)

RESUMEN

The roles of O-acetylserine (thiol) lyase (OASTL, EC 4.2.99.8) and abscisic (ABA) acid in stress responses to NaCl and cadmium treatments were investigated in Typha latifolia L. and Phragmites australis (Cav.) Trin. ex Steudel plants. OASTL activity increased under stress (25-300 microM Cd, 100mM NaCl, 1 microM ABA) in both Typha and Phragmites mainly in roots, contributing substantially to satisfy the higher demand of cysteine for adaptation and protection. The earliest significant responses in intact roots were recorded after 12-24 h of Cd treatments, but different levels of stimulation were also observed after 3 and 7 days of exposure. The OASTL activity responses of Phragmites to salinity (100mM NaCl) were higher than those of Typha. Cysteine synthesis in Typha is much higher than in Phragmites, which supports the efficiency of the thiol-metabolism-based protection shown in Typha. Exogenous ABA increased OASTL activity in both species. Cd treatments led to increased ABA levels in roots. Phragmites showed higher ABA levels compared to Typha. The increase of ABA content indicates the involvement of this phytohormone in early stress responses, while the stimulation of OASTL following the ABA application suggests that ABA has a role in an OASTL activation pathway.

Asunto(s)

RESUMEN

ABA is a plant signalling-molecule that plays a key role in regulating stomatal response, stress-tolerance responses, and coordinated growth of roots and shoots. Knowledge of the relationship between endogenous ABA level and plant growth is essential for improving plant growth and productivity. The wild-type tomato Rheinlands Ruhm (RR) and its flacca mutant were grafted in order to determine the relationship between endogenous ABA levels and biomass production. The flacca genotype is an ABA-deficient mutant, characterized by high stomatal conductance during the day or the night, high transpiration rate, marked tendency to wilt, and smaller size. Flacca scions grafted on to wild-type rootstock (Fs/Wr) exhibited higher ABA levels, lower transpiration rate, and higher water content than those of a control graft of flacca scion on flacca rootstock (Fs/Fr). Fs/Wr exhibited a lower ABA concentration, xylem exudate rate, ABA xylem-loading rate, and dry weight biomass in wild-type rootstock than in control grafts of wild-type scion on wild-type rootstock (Ws/Wr). Flacca rootstock grafted to wild-type scion (Ws/Fr) showed a higher ABA level, xylem exudation rate, ABA xylem-loading rate, dry weight biomass and length than grafts to flacca scion (Fs/Fr). Ws/Fr did not induce significant changes in wild-type scion as compared with Ws/Wr. In double grafts, flacca shoot fresh weight was significantly increased in flacca scion and wild-type scion grafted on to flacca rootstock (Fs + Ws/Fr) or wild-type rootstock (Fs + Ws/Wr). There was a significant linear relationship between biomass and ABA in scions (r=0.997, P=0.001). These results support the notion that ABA increases growth of tomato seedlings via improved stomatal control.

RESUMEN

Pea plants (Pisum sativum L.) grown initially in nutrient solutions with adequate nitrogen supply (4 mM NO3-) were transferred to solutions containing salt (50 or 100 mM NaCl), ammonium (4 mM) or a low nitrogen supply (0.4 mM NO3-). No changes of abscisic acid (ABA) content were found in roots of stressed pea plants 9 d after the beginning of the treatments; however, accumulation of ABA in the leaves was observed. Old leaves accumulated ABA to a higher extent than young leaves. Accumulation of ABA in leaves of ammonium-fed plants and plants grown under low nitrogen supply occurred in the absence of both increased ABA xylem loading rate and enhanced aldehyde oxidase (AO, EC 1.2.3.1) activity in roots. Enhanced leaf AO activity was observed in all treatments, with the highest increase in old leaves. Among the three AO isoforms (AO-1, AO-2 and AO-3) detected in extracts of pea leaves, the lowest one AO-3 (highest mobility in the gel) correlated with ABA production and showed the highest increment in response to the treatments. The increase of AO activity detected in leaves after 2 weeks of stress application was less prominent than after 9 d, suggesting a transient enhancement of ABA production following the onset of stress. An increase of ABA xylem loading rate as well as AO root activity 4 d and 9 d after application of the treatments was observed only in salt-treated plants followed by a decrease after 14 d in 100 mM NaCl. Decreased cytokinin (trans-zeatin riboside) delivery rate into the xylem sap was observed in all treatments. The role of abscisic acid and cytokinins as positive and negative growth signals, as well as the involvement of root-generated ABA on ABA accumulation in leaves is discussed.

Asunto(s)

RESUMEN

The variant surface glycoprotein (VSG) genes of Trypanosoma brucei are transcribed in telomeric loci termed VSG expression sites (ESs). Despite permanent initiation of transcription in most if not all of these multiple loci, RNA elongation is abortive except in bloodstream forms where full transcription up to the VSG occurs only in a single ES at a time. The ESs active in bloodstream forms are polycistronic and contain several genes in addition to the VSG, named ES-associated genes (ESAGs). So far 12 ESAGs have been identified, some of which are present only in some ESs. Most of these genes encode surface proteins and this list includes different glycosyl phosphatidyl inositol (GPI)-anchored proteins such as the heterodimeric receptor for the host transferrin (ESAG7/6), integral membrane proteins such as the receptor-like transmembrane adenylyl cyclase (ESAG4) and a surface transporter (ESAG10). An interesting exception is ESAG8, which may encode a cell cycle regulator involved in the differentiation of long slender into short stumpy bloodstream forms. Several ESAGs belong to multigene families including pseudogenes and members transcribed out of the ESs, named genes related to ESAGs (GRESAGs). However, some ESAGs (7, 6 and 8) appear to be restricted to the ESs. Most of these genes can be deleted from the active ES without apparently affecting the phenotype of bloodstream form trypanosomes, probably either due to the expression of ESAGs from 'inactive' ESs (ESAG7/6) or due to the expression of GRESAGs (in particular, GRESAGs4 and GRESAGs1). At least three ESAGs (ESAG7, ESAG6 and SRA) share the evolutionary origin of VSGs. The presence of these latter genes in ESs may confer an increased capacity of the parasite for adaptation to various mammalian hosts, as suggested in the case of ESAG7/6 and proven for SRA, which allows T. brucei to infect humans. Similarly, the existence of a collection of slightly different ESAG4s in the multiple ESs might provide the parasite with adenylyl cyclase isoforms that may regulate growth in response to different environmental conditions. The high transcription rate and high recombination level that prevail in VSG ESs may have favored the generation and/or recruitment in these sites of genes whose hyper-evolution allows adaptation to a larger variety of hosts.

Asunto(s)

RESUMEN

The distribution of the Mo-enzymes aldehyde oxidase (AO; EC 1.2.3.1) xanthine dehydrogenase (XDH; EC 1.2.1.37) and nitrate reductase (NAD(P)H NR; EC 1.6.6.1-2) was studied along the longitudinal and transversal axes of maize (Zea mays L. cv. Jubily) nodal roots as affected by nitrogen sources and salinity. Activities of the Mo-enzymes were considerably enhanced under mild saline conditions. The activities of AO and XDH increased following addition of ammonium to the nutrient solution. Immunoblot analysis with antibodies raised against maize AO protein revealed increased levels of AO proteins in root tips of ammonium fed plants. Application of salinity to nitrate fed plants did not affect the enzyme protein level, although it enhanced the activity of the Mo-hydroxylases. The specific activities of the Mo-enzymes were the highest in root tips (0-1 cm segments) while on the transversal axis maximal activity was observed in the stele or vascular cylinder. Activity staining of AO after native PAGE of root extracts revealed four bands of AO proteins (AO1-4) capable of oxidizing a number of aliphatic and aromatic aldehydes. Increased AO activity in maize nodal roots grown with ammonium, and salinity were observed mainly at the AO3 and AO4 bands. Tips and stele contained primarily AO3 and AO4, and only traces of AO1 and AO2. SDS-PAGE of root extracts followed by Western blots revealed, besides the major 150 kD subunit of AO, two polypeptides with molecular masses of 72 and 85 kD located specifically in the cortex. Part of the polymorphism of AO in plant roots may be related to the allocation of distinct isoforms to different regions of the root, although the specific metabolic roles of the different bands have not been established.

RESUMEN

In Trypanosoma brucei the genes are organised into long polycistronic transcription units and only three promoters for protein-encoding genes and a single terminator have been characterised. These promoters recruit a polI-like RNA polymerase for the transcription units encoding the two major stage-specific antigens of the parasite, the variant surface glycoprotein (VSG) of the bloodstream form and procyclin of the insect-specific procyclic form, while the terminator is that of a procyclin transcription unit. By deletional and mutational analysis we defined the two DNA sequences essential for the activity of the VSG promoter from a bloodstream form transcription unit and one of the functional elements of the procyclin terminator. These three short sequences are similar, and their C-rich strand binds the same protein of 40 kDa. In addition, this factor also binds to the C-rich strand of the telomeric repeats, the consensus target sequence being 5'-CCCTNN-3'. The factor-binding sequences are functionally interchangeable in chimeric promoter or terminator constructs, although additional elements are required for full activity.

Asunto(s)

Asunto(s)

RESUMEN

The polycistronic procylcin PARP (for procyclic acidic repetitive protein) A transcription unit of Trypanosoma brucei was completely characterized by the mapping of the termination region. In addition to the tandem of procyclin genes and GRESAG 2.1, this 7.5- to 9.5-kb unit contained another gene for a putative surface protein, termed PAG (for procyclin-associated gene) 3. The terminal 3-kb sequence did not contain significant open reading frames and cross-hybridized with the beginning of one or several transcription units specific to the bloodstream form. At least three separate fragments from the terminal region were able to inhibit chloramphenicol acetyltransferase expression when inserted between either the PARP, the ribosomal, or the variable surface glycoprotein promoter and a chloramphenicol acetyltransferase reporter gene. This inhibition was due to an orientation-dependent transcription termination caused by the combination of several attenuator elements with no obvious sequence conservation. The procyclin transcription terminator appeared unable to inhibit transcription by polymerase II.

Asunto(s)

RESUMEN

PURPOSE: This phase I study of Taxotere (RP 56976, NSC 628503; docetaxel, Rhône-Poulenc Rorer, Antony, France) was undertaken to determine the maximum-tolerated dose (MTD), toxic effects, and basic pharmacokinetics of a day-1 and -8 schedule of this novel semisynthetic product related to Taxol (paclitaxel; Bristol-Myers Squibb, Wallingford, CT). PATIENTS AND METHODS: Thirty-two eligible patients with refractory solid malignancies have been treated with a 1-hour infusion of Taxotere on a day-1 and -8 schedule every 3 weeks as long as patients maintained a polymorphonucleotide count > or = 1,500/microL and a platelet count > or = 100,000/microL. Dose levels tested have ranged between 20 and 110 mg/m2 per course. RESULTS: Considering 128 assessable courses, the main toxicities have been neutropenia (which was dose-limiting), asthenia, alopecia, hypersensitivity reactions, skin toxicity, and edema. No significant cardiac or platelet toxicity has been observed. Seven patients have had aggravation of preexisting paresthesias or new onset of sensory symptoms during Taxotere treatment. The MTD at this schedule appears to be 110 mg/m2 per course, with six of 10 patients at this level experiencing severe toxicity. Five partial remissions have been observed in four heavily pretreated patients with breast cancer and in one patient with adenocarcinoma of unknown origin. Two patients with ovarian cancer have had meaningful decreases in CA125 levels. CONCLUSION: Like Taxol, this novel chemotherapeutic agent appears to possess promising activity in patients with refractory breast and ovarian neoplasms, with tolerable toxicities. Using this schedule, 100 mg/m2 per course is the recommended dose for future phase II trials.

Asunto(s)

Asunto(s)

RESUMEN

The P-type ATPase gene TBA1 of Trypanosoma brucei belongs to a polycistronic transcription unit. We analyzed the structure and expression of a 4-kb region located immediately downstream from TBA1. This region is unique and contains two large open reading frames transcribed into stable mRNAs. These putative genes, termed ADG1 and ADG2, can respectively encode a 24-kDa and a 81-kDa protein. The intergenic spacings between the polyadenylation sites and the next 3' splice acceptor sites are very short: 148 bp between TBA1 and ADG1, and 127 bp between ADG1 and ADG2. Transcripts from each of the two ADG1 alleles can be detected, indicating that both homologs are transcribed. These transcripts are differentially spliced due to a single base difference which destroys in one homolog the AG acceptor site present in the other. In the 'mutant' allele an alternative downstream splice acceptor site is used. Despite its sequence conservation in both alleles, this splice site is only used in the allele lacking the upstream AG acceptor site. The major population of ADG1 transcripts exhibit a long 5'-untranslated extension and no 3'-terminal tail, but a minor population shows a smaller 5'-untranslated region due alternative splicing closer to the initiation codon of the gene. The steady-state amounts of transcripts from individual genes in this region are differentially stage-regulated.

Asunto(s)

RESUMEN

Clebopride, a new benzamide derivative, has, in common with the other members of this group, antidopaminergic activity. In animals, its therapeutic ratio is superior to that of metoclopramide at doses free of side effects associated with hyperprolactinemia and extrapyramidal symptoms. The present study was designed to define the maximum tolerated dose (MTD) in patients with advanced histologically-proven cancer, treated with cisplatin at a dose of greater than 50 mg/m2. Most of them were pretreated and refractory to standard antiemetics. Clebopride was started at a dosage of 0.10 mg/kg in a group of 6 patients and escalated by 0.2 mg at each dose level. A total of 30 patients were included. Side effects include somnolence, diarrhea and extrapyramidal-like symptoms. The latter occurred at almost all dose levels in 14% of the cycles and limited continuation of the study. Activity in this group of patients was encouraging but, considering the rate of extrapyramidal symptoms, further dose escalation is not indicated and activity at lower, nontoxic levels should be investigated.

Asunto(s)

RESUMEN

The transcription unit of the gene for the variant specific glycoprotein (VSG) AnTat 1.3A of Trypanosoma brucei contains several associated genes (ESAGs, for Expression Site-Associated Genes), 7 of which have already been described. We report here the characterization of a further ESAG, which we term ESAG 8, present 1 kb downstream from the putative adenylate cyclase gene ESAG 4. ESAG 8 encodes a 70 kd protein whose sequence indicates that it is probably not exposed at the cell surface. With the exception of the N-terminal domain which contains a presumptive DNA-binding zinc finger, the ESAG 8 protein consists exclusively of leucine-rich repeats of 23 amino acids, typical of protein-interacting domains such as the RAS-interacting region of the yeast adenylate cyclase. ESAG 8 transcripts are only found in bloodstream forms, and their level is particularly low, suggesting a high rate of degradation. The ESAG 8 protein may be involved in stage-specific regulatory processes, such as gene expression control or adenylate cyclase activation.

Asunto(s)

RESUMEN

The effect of ammonium and nitrate nutrition on maize and wheat grown hydroponically and salinity stressed was assessed from measurements of growth rate and gas exchange. In both maize and wheat the ammonium-grown plains were much more sensitive to salinity toxicity than nitrate-grown plants particularly when exposed to 60-80 mM salinity. Shoot growth was retarded to a far greater extent than root growth in salinity-stressed plants of both wheat and maize with either nitrogen source. There was no significant decrease of photosynthetic rate in salinity-stressed plants of either species fed nitrate or ammonium, except in severely wilted plants of both species fed nitrate or ammonium at the highest (80 mM) salinity concentration. The same was true for stomatal conductance, transpiration rate and transpiration ratio (water use efficiency). In nitrate-fed wheat, raising the calcium concentration from 2 to 12 mM in the presence of 60 mM salinity produced an 11% increase in growth. This effect is ascribed to improved nitrate uptake due to calcium protection of the nitrate transporter and was not evident in ammonium-grown wheat. Possible reasons for the differential effects of ammonium and nitrate nutrition are discussed.

RESUMEN

Glutaraldehyde fixation in 0.33 M sorbitol without any buffer reveals changes in the staining properties of the envelopes of chloroplasts of pea plants kept in the light or in the dark prior to fixation. After dark pretreatment the outer double membrane of the chloroplast does not adsorb heavy metals, resulting in a "white" unstained rim instead of the usual membrane. All other membranes of the cell, including chloroplast grana, are not affected and stain normally. Light pretreatment of the plants allows the usual staining of the outer membrane of the chloroplats. Fixation carried out in the medium usually used to isolate intact CO2 fixing chloroplasts (sorbitol+buffer+ions) reverses the above process and results in unstained envelopes of chloroplasts from preilluminated leaves, while the envelopes of chloroplasts from leaves kept in the dark stain normally. Glutaraldehyde-fixed chloroplats isolated from preilluminated leaves show a very basic isoelectric point during electrofocusing, while fixed chloroplasts from predarkened tissue exhibit an isoelectric point at about pH 7.

RESUMEN

Comparative studies of nitrate-activated nitrate reductase (NR-NO2) and nitrate-induced nitrate reductase (NR-NO3) (EC 1.6.6.2) indicate that the enzymes differ in structure, heat stability, and pH dependence, but have the same cofactor requirment. NR-NO2 developes in barley (Hordeum vulgare L. var. Dvir) seedlings as NR-NO3 disappears. A transition from the active to the inactive form of nitrate reductase takes place. Nitrite seems to activate the inactive form of the enzyme.

RESUMEN

Homogenates of dark-pretreated leaves yield two particulate fractions in density gradient centrifugation: one contains chlorophyll (chloroplasts) while a second fraction contains ribulose-1, 5-bisphophate carboxylase, NADP-dependent glyceraldehyde-3-phosphate dehydrogenase and catalase. Addition of a microbody-rich pellet to chloroplasts isolated from dark-pretreated plants largely enhances both oxygen evolution and CO2-fixation into organic compounds. The pathway of CO2 reduction may be part of a membrane system which, under suitable conditions, may separate from the chloroplast as a distinct cytoplasmic entity, having physical properties similar to those of microbodies.

Asunto(s)

RESUMEN

The content of specific enzymes in microbodies isolated from tobacco Nicotiana rustica, L. leaves may vary according to the procedure followed during the isolation of the organelles. The type of homogenizing medium, its ionic components and the ratio of medium to tissue during homogenization, affect the over-all yield and relative distribution of each microbody enzyme in different ways. The type of density gradient and the initial acceleration of the centrifuge rotor affect also the enzyme content of sedimenting microbodies. These observations explain some of the conflicting results obtained on the determination of the intracellular location of several enzymes.

RESUMEN

Glycolate oxidase is loosely held by microbodies obtained from etiolated barley (Hordeum vulgare L.) leaves depleted of nitrate. Defined centrifugation conditions cause the complete detachment of the enzyme from the microbodies. Addition of nitrate to these plants brings about a greater retention of glycolate oxidase by the microbodies. Synthesis of a nitrate-induced protein seems to be responsible for the enhanced retention of glycolate oxidase. Catalase, on the contrary, is strongly attached to the microbodies under all nutritional and experimental conditions considered.