A core outcome set for future endometriosis research: an international consensus development study.

OBJECTIVE: To develop a core outcome set for endometriosis. DESIGN: Consensus development study. SETTING: International. POPULATION: One hundred and sixteen healthcare professionals, 31 researchers and 206 patient representatives. METHODS: Modified Delphi method and modified nominal group technique. RESULTS: The final core outcome set includes three core outcomes for trials evaluating potential treatments for pain and other symptoms associated with endometriosis: overall pain; improvement in the most troublesome symptom; and quality of life. In addition, eight core outcomes for trials evaluating potential treatments for infertility associated with endometriosis were identified: viable intrauterine pregnancy confirmed by ultrasound; pregnancy loss, including ectopic pregnancy, miscarriage, stillbirth and termination of pregnancy; live birth; time to pregnancy leading to live birth; gestational age at delivery; birthweight; neonatal mortality; and major congenital abnormalities. Two core outcomes applicable to all trials were also identified: adverse events and patient satisfaction with treatment. CONCLUSIONS: Using robust consensus science methods, healthcare professionals, researchers and women with endometriosis have developed a core outcome set to standardise outcome selection, collection and reporting across future randomised controlled trials and systematic reviews evaluating potential treatments for endometriosis. TWEETABLE ABSTRACT: @coreoutcomes for future #endometriosis research have been developed @jamesmnduffy.

Transforming growth factor-ß1 functional polymorphisms in myeloablative sibling hematopoietic stem cell transplantation.

Hematopoietic stem cell transplantation (HSCT) with sibling donors (s.d.) is a life-saving intervention for patients with hematological malignancies. Numerous genetic factors have a role in transplant outcome. Several functional polymorphisms have been identified in TGF-ß1 gene, such as single-nucleotide polymorphism (SNP) at +29C>T within exon 1. Two hundred and forty five patient/donor pairs who underwent a s.d. HSCT in our centers were genotyped for this SNP. In the myeloablative cohort, +29CC donors were associated with an increase in severe chronic GvHD (32% vs 16%, hazard ratio (HR) 9.0, P=0.02). Regarding survival outcomes, +29CC patients developed higher non relapse mortality (NRM) (1-5 years CC 28-32% vs TC/TT 7-10%; HR 5.1, P=0.01). Recipients of +29TT donors experienced a higher relapse rate (1-5 years TT 37-51% vs TC 19-25% vs CC 13%-19%; HR 2.4, P=0.01) with a decreased overall survival (OS) (1-5 years TT 69-50% vs TC/CC 77-69%; HR 1.9, P=0.05). Similar to previous myeloablative unrelated donors HSCT results, we confirmed that +29CC patients had higher NRM. In addition we found that +29TT donors might be associated with a higher relapse rate and lower OS. These results should be confirmed in larger series. Identification of these SNPs will allow personalizing transplant conditioning and immunosuppressant regimens, as well as assisting in the choice of the most appropriate donor.

Root Responses to Boron Deficiency Mediated by Ethylene.

Low boron (B) supply alters the architecture of the root system in Arabidopsis thaliana seedlings, leading to a reduction in the primary root growth and an increase in the length and number of root hairs. At short-term (hours), B deficiency causes a decrease in the cell elongation of the primary root, resulting in a lower growth. Experimental approaches using ethylene insensitive Arabidopsis mutants, inhibitors of ethylene response, and GUS reporter lines suggest that ethylene is involved in these responses of the primary root to B deficiency. Furthermore, it has been shown that auxin participates in the inhibition of cell elongation under short-term B deprivation. These results support that an interaction between ethylene and auxin plays an important role in controlling the primary root elongation, in which a number of genes related to the synthesis, transport, and signaling of both phytohormones could modulate this effect. Evidence for a root cross-talk among both hormones and other possible intermediates (abscisic acid, calcium sensors, and reactive oxygen species) in response to B deficiency is provided and discussed.

The effect of carbon dioxide on the shelf life of ready-to-eat shredded chicken breast stored under refrigeration.

The objective of the present study was to determine the shelf life of ready-to-eat cooked chicken breast fillets (shredded) stored in atmospheres that were modified with different concentrations of CO2 and to establish a relationship between the concentration of this gas and bacterial growth. The samples were divided into 7 groups with different packaging conditions: aerobiosis, vacuum, and 10, 30, 50, 70, and 90% CO2 (with the remaining volume filled with N2). All of the samples were stored at 4 ± 2°C for 28 d. During this period, pH tests and counts of aerobic heterotrophic mesophyll bacteria (AHMB), aerobic heterotrophic psychotropic bacteria (AHPB), Enterobacteriaceae, and lactic acid bacteria (LAB) were performed, and the gas compositions of the packaging atmospheres were verified. The pH of the aerobic packages increased during storage. However, the other treatments resulted in the opposite trend, with the CO2 concentration decreasing over the first 24 h and then remaining constant until the end of experiment. A gradual increase in the AHMB, AHPB, Enterobacteriaceae, and LAB counts was observed during storage; this increase was faster in the meat that was packed under aerobiosis conditions than in the other treatments. The treatments with a CO2 concentration above 10% exhibited lower Enterobacteriaceae growth, whereas LAB growth was discrete in all of the treatments, independent of the CO2 concentration. The shelf life of the samples packed with 90% CO2 was 28 d. Based on the AHMB and AHPB counts, the shelf life was 3 times longer than for the samples packed under aerobiosis conditions (9 d). The increased package CO2 concentration caused a reduction in the growth rate of the examined bacteria (r = 0.99), and treatment with 90% CO2 appears promising as a method with which to increase the product's shelf life.

Stimulatory effect of PGF2α on PRL based on experimental inhibition of each hormone in mares.

During the luteolytic period in mares, the peak of 65% of pulses of a PGF2α metabolite (PGFM) and the peak of a pulse of PRL have been reported to occur at the same hour. It is unknown whether the synchrony reflects an effect of PGF2α on PRL or vice versa. Controls, a flunixin meglumine (FM)-treated group (to inhibit PGF2α), and a bromocriptine-treated group (to inhibit PRL), were used at 14 days postovulation in June and in September (n = 6 mares/group/mo). Blood samples were collected hourly from just before treatment (Hour 0) to Hour 10. Concentrations of PGFM in the FM group were lower (P < 0.05) at Hours 4 to 6 than in the controls in each month, but bromocriptine had no detected effects on PGFM. Concentrations of PGFM averaged over all groups and within each group did not differ between June and September. Compared to the controls, concentrations of PRL in June were lower (P < 0.05) in the FM group at Hours 4 to 8 and in the bromocriptine group at Hours 4 to 10. Concentration of PRL averaged over groups was lower (P < 0.0001) in September (0.9 ± 0.05 ng/mL, mean ± SEM) than in June (3.0 ± 0.3 ng/mL). Results supported the hypothesis that the positive association between PGFM and PRL concentrations in mares represents an effect of PGF2α on PRL rather than an effect of PRL on PGF2α.

Role of LH in luteolysis and growth of the ovulatory follicle and estradiol regulation of LH secretion in heifers.

The role of LH in luteolysis and development of the ovulatory follicle and the involvement of GnRH receptors in estradiol (E2) stimulation of LH secretion were studied in heifers. A pulse of PGF(2α), as indicated by a metabolite, was induced by E2 treatment on Day 15 (Day 0 = ovulation) and LH concentration was reduced with a GnRH-receptor antagonist (acyline) on Days 15, 16, and 17. Blood samples were collected every 6 h on Days 14-17 and hourly for 10 h beginning at the Day-15 treatments. Four groups were used (n = 6): control, acyline, E2, and E2/acyline. The number of LH pulses/heifer during the 10 h posttreatment was greater (P < 0.0002) in the E2 group (2.3 ± 0.4, mean ± SEM) than in the acyline group (0.2 ± 0.2) and was intermediate in the E2/acyline group (1.4 ± 0.2). Concentrations of progesterone in samples collected every 6 h on Day 15 showed a group-by-hour interaction (P < 0.02); concentrations decreased in the acyline group but not in the control group. The 12 heifers in the combined acyline and E2/acyline groups had three follicular waves compared to two waves in 10 of 12 heifers in the combined control and E2 groups. Results (1) supported the hypothesis that LH delays the progesterone decrease associated with luteolysis, (2) supported the hypothesis that LH has a positive effect on the continued development and growth of the selected ovulatory follicle, and (3) indicated that E2 stimulates LH production through an intracellular pathway that involves GnRH receptors on the gonadotropes and a pathway that does not involve the receptors.

Concentrations of circulating hormones during the interval between pulses of a PGF2α metabolite in mares and heifers.

The temporal relationship of several hormones to a metabolite of prostaglandin F2α (PGFM) was studied in mares and heifers from the beginning of the first PGFM pulse during luteolysis to the end of the second pulse. Mares (n=7) were selected with a 9-h interval between the peaks of the two pulses. In mares, estradiol-17ß (estradiol) increased (P<0.05) within each PGFM pulse and plateaued for a mean of 6h between the pulses, resulting in a stepwise estradiol increase. Progesterone decreased linearly (P<0.0001) throughout the intra-pulse and inter-pulse intervals of PGFM. In heifers (n=6), inter-pulse intervals were variable, and therefore Hours 1-4 of the first pulse (Hour 0=PGFM peak) and Hours -4 to -1 of the second pulse were used to represent the mean 8-h interval between peaks of the two pulses. Estradiol increased (P<0.05) during the ascending portion of each PGFM pulse and then decreased (P<0.05) beginning at Hour -1 of the first PGFM pulse and Hour 0 of the second pulse. The 1-h delay during the second pulse was accompanied by an apparent increase in PRL. A transient decrease in estradiol occurred in individuals between PGFM pulses at a mean of 5h after the first PGFM peak, concomitant with a transient LH increase (P<0.05). Results indicated that estradiol plateaued in mares and fluctuated in heifers during the interval between PGFM pulses. Heifers also showed temporal relationships between estradiol and LH and apparently between estradiol and PRL.

[Morphologic changes in the proximal jejunum after partial hepatectomy in rats]. / Estudio de los cambios morfológicos en el yeyuno proximal después de hepatectomía parcial en la rata.

INTRODUCTION: Partial hepatectomy is being used with increasing frequency and wider surgical indications including living donor transplantation, particularly in children. Our aim was to study the morphologic changes that occur in the proximal jejunum as a result of the anatomic, functional, and metabolic unity of the liver and small intestine. MATERIAL AND METHODS: Twenty-four Wistar rats were divided into 2 groups, a control group and an experimental group (30, 90, and 180 days). Changes in the proximal jejunum after 70% partial liver resection were studied by evaluating 4 parameters (total wall width, villous height, crypt depth, and villous thickness). RESULTS: Jejunal total wall width increased significantly in the experimental group (p < 0.001). Villous height increased significantly in the experimental group as compared to the control group (p < 0.001). Villous thickness increased, and crypt depth decreased in the experimental group, but these changes were not statistically significant. CONCLUSIONS: Partial hepatectomy (70%) originated trophic changes in the proximal jejunum that persisted in the short and long term, giving rise to parietal and villous hypertrophy.

A simple procedure to perform intravenous injections in the Mongolian gerbil (Meriones unguiculatus).

We describe a simple and feasible procedure for performing intravenous administration of substances in the gerbil. Under light anaesthesia, animals were held in dorsal recumbency and a very small incision of skin, parallel to the femoral vein on the internal side of the thigh, was made. The vein is easily accessible via thin skin incision. An insulin syringe and a 30 G needle were used for the injection. This is an easy and quick method, which, with appropriate anaesthesia, allows rapid recovery.

Evaluation of alternative reporter genes for the yeast two-hybrid system

The yeast two-hybrid system is a powerful tool for screening protein-protein interactions and has also been used for large-scale studies. We evaluated two protein-coding sequences as reporter genes for the yeast two-hybrid system, to determine if it was suitable as an alternative screening strategy. Aspergillus awamori glucoamylase activity results in clear haloes around colonies producing this enzyme after growth on starch plates and staining with iodine vapors. However, transcription activation by Gal4 on Gal-regulated promoters was insufficient for this type of phenotypic visualization. A modified gene of Aequoria victoria enhanced green fluorescent protein (EGFP) was tested to determine its suitability for interaction screenings with flow cytometry. When the EGFP reporter gene system was incorporated into the cells, Gal4 transcriptional activation produced sufficient fluorescence for detection with the flow cytometer, especially when there were strong interactions

[Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero]. / Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero.

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The "in house" indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.

Ausencia de circulación de los virus HTLV-I/II en donantes de sangre de las provincias de Santa Fe y Santiago del Estero / Absence of HTLV-I/II virus circulation in blood donors from the provinces of Santa Fe and Santiago del Estero

In the present study, we evaluated the seroprevalence of HTLV-I/II infection among the blood donors in Santa Fe and Santiago del Estero provinces. A total of 1327 serum samples from blood donors from Rafaela blood bank of Santa Fe province and 3382 serum samples from blood donors from Dr Edgar Bouzon blood bank of Santiago del Estero province were studied. The antibody screening was done by particle agglutination assay (PA) (SERODIA, Fujirebio Inc., Tokyo, Japan) or by enzyme immuno assay (EIA) (Abbott HTLV-I/HTLV-II EIA, Abbott, Germany). The "in house" indirect immunofluorescence assay (IFA) and Western blot (Bioblot HTLV Biokit, Barcelona, Spain) were used as confirmatory assays. All the samples resulted negative for specific antibodies against HTLV-I/II. These results suggest that HTLV-I/II are not circulating in low risk populations in these provinces or that the prevalences of infection would be lower than that reported by blood banks in other provinces of Argentina.

Diagnosis of renal agenesis in a beagle.

A case of right renal agenesis in a beagle, of interest because of the age of the dog at the time of diagnosis, is described. Physical, haematological, biochemical and urinary examinations, including measurement of endogenous creatinine clearance, were performed to assess renal function. Survey radiography, excretory urography, ultrasonography, computed tomography and nuclear magnetic resonance examinations were also used to confirm the absence of a kidney. The effect of kidney agenesis on renal function, evaluated on the basis of endogenous creatinine clearance, is discussed together with the benefits of the various imaging techniques to enable in vivo detection of renal abnormalities.

Comparison of chagasic and non-chagasic myocardiopathies by ELISA and immunoblotting with antigens of Trypanosoma cruzi and Trypanosoma rangeli.

Trypanosoma cruzi associated myocardiopathy, or Chagas disease, continues to be a serious problem in Venezuela, for which there is neither a vaccine nor a cure. In order to learn more about the humoral immune response to trypanosomal antigens, and to try to identify dominant antigens, we used ELISA and immunoblotting to study the reactivity of sera from patients with chagasic and non-chagasic myocardiopathies, against surface and secreted proteins from T. cruzi and T. rangeli. Both species are found in the same insect vector, but only T. cruzi is thought to be pathogenic in vertebrates. The ELISA results fell into three patterns: (1) high reactivity values with both T. cruzi and T. rangeli surface and secreted proteins; (2) high values to T. cruzi but low values with T. rangeli; and (3) high values to T. rangeli and low values with T. cruzi. This finding that some chagasic sera react more strongly against T. rangeli than against T. cruzi is intriguing, and warrants further investigation. When chagasic sera were tested on Western blots of total extracts of T. cruzi and T. rangeli, the pattern of reactive bands was similar against both parasites, but no two sera showed an identical pattern. Furthermore, there was no correlation between a particular immunoblotting pattern and either the antibody titer, or the severity of the disease. Several T. cruzi and T. rangeli antigens were recognized by sera from healthy controls as well as from patients with other tropical diseases endemic in Venezuela. Overall, our results suggest that the humoral immune response to trypanosomal antigens is complex, and no single antigen may be the determining factor in the pathogenesis of chagasic myocardiopathy.

Perception of sweetness and bitterness in different vehicles.

In the present study, we investigated taste-taste, taste-vehicle, and simultaneous taste-vehicle-taste mixtures. Subjects made estimates of the sweetness and bitterness of 27 stimuli. Sucrose (292, 585, and 1170 mM), caffeine (13, 26, and 52 mM), and binary mixtures of low (292-13 mM), middle (585-26 mM), and high (1170-52 mM) levels of both components were dispersed in water, carboxymethylcellulose (CMC) 1% w/v, and gelatin 6% w/v. The sweetness and bitterness of the sucrose-vehicle-caffeine combinations were significantly weaker than the respective sucrose-vehicle and caffeine-vehicle combinations. The emerged mutual suppressive effects were asymmetrical and persisted when both tastants were presented in CMC and gelatin. Moreover, the increase in vehicle consistency and the simultaneous addition of another taste reduced the perceived intensity of a taste either presented alone or dissolved in water. For both sweetness and bitterness, the total taste suppression observed was always significant.

Glycation of blood proteins during pregnancy and lactation in the rat.

Non-enzymatic glycation of blood proteins is a time and concentration dependent process and has been used clinically to monitor carbohydrate metabolism during human pregnancy. Since gestation in rats is of much shorter duration than in humans (3 weeks vs 9 mos) the question was raised whether similar differences in glycated proteins could be observed. Therefore, levels of glucose, glycated hemoglobin and fructosamine were measured during normal pregnancy and lactation in rats. Glucose levels during late pregnancy were significantly lower than in non pregnant and early pregnant rats. During lactation glucose levels return to normal. Glycated hemoglobin paralleled glucose decrease during late pregnancy and increased during lactation. Fructosamine followed a similar pattern. Therefore glycated hemoglobin and fructosamine appear to be reliable indicators of glucose status during gestation and lactation similar to humans and may have value as predictors of gestational diabetes mellitus once a suitable rat model is developed.

Differential growth requirements of several Leishmania spp. in chemically defined culture media.

17 strains of Leishmania from 4 species: brasiliensis, mexicana, donovani and garnhami have been continually cultured at 26 degrees C, in the absence of proteins, in a medium containing salts, glucose, D-ribose, 2-deoxyribose, hemin, tricine, HEPES, 34 amino acids and intermediates of amino acid metabolism, 23 vitamins, 6 nucleotides and tetrahydrofolic acid. A wide variation in growth requirements was observed among leishmaniae which permitted the preparation of different minimum culture media for each Leishmania spp. Virulence of parasites was maintained after 30 passages in these chemically defined media. The requirements for differentiation to amastigotes also varied among the species as a function of the temperature of incubation and the protein content of the culture medium. Bovine serum albumin tryptic peptides substituted fetal bovine serum as growth factors at 30-34 degrees C.

Trypanosoma cruzi: growth requirements at different temperature in fetal bovine serum or peptide supplemented media.

An enriched synthetic medium with low molecular weight peptides allows Trypanosoma cruzi epimastigotes to grow at 26-37 C. Using this medium, the growth requirements of T. cruzi were compared at different temperatures. When supplemented with fetal bovine serum or serum peptides, nine amino acids were absolutely required from the first passage, while additional amino acids and amino acid precursors were needed to support growth during a second passage. Five amino acids (beta-alanine, glutamine, cysteine, ornithine, and threonine) were also required absolutely at temperatures ranging between 30 and 37 C. Nine vitamins were needed at all temperatures, while ascorbic acid and ergocalciferol were not necessary at any temperature. The remaining amino acids and vitamins showed a variable role as growth factors depending on the temperature increase. In peptide supplemented media, requirements for amino acids and their precursors, as well as vitamins and nucleotides, increased markedly when compared with the protein supplemented medium. A peptide composed of one glutamic acid, two alanines, and one lysine can substitute for serum for trypanosomal growth at all temperatures. Several minimum media have been prepared in which epimastigote forms of T. cruzi can grow at 26-37 C for more than 10 passages.

Protein and nucleotide contamination of bovine liver catalase used in culture medium explains growth of Trypanosoma cruzi.

Commercially available bovine liver catalase has been used to supplement chemically defined medium for growth of Trypanosoma cruzi. The protein extract was found to be contaminated with 25 to 30 protein bands as well as DNA and RNA polymers.