Search

Background: Non-compressible abdominal hemorrhage (NCAH) is the leading cause of potentially preventable deaths in both civilian and military austere environments, and an improvement in mortality due to this problem has not been demonstrated during the past quarter century. Several innovations have been developed to control hemorrhage closer to the point of injury., Objective: This review assessed NCAH interventions in civilian and military settings, focusing on austere environments. It identified innovations, effectiveness, and knowledge gaps for future research., Methodology: The Joanna Briggs Institute for Evidence Synthesis methodology guided this scoping review to completion. Studies evaluating NCAH with human participants in civilian and military austere environments that were eligible for inclusion were limited to English language studies published between December 1990 and January 2023. The PCC (Participant, Concept, Context) framework was used for data synthesis. Deductive and inductive thematic analyses were used to assess the literature that met inclusion criteria, identify patterns/themes to address the research questions and identify common themes within the literature. A stakeholder consultation was conducted to review and provide expert perspectives and opinions on the results of the deductive and inductive thematic analyses., Results: The literature search identified 868 articles; 26 articles met the inclusion criteria. Textual narrative analysis of the 26 articles resulted in the literature addressing four main categories: NCAH, penetrating abdominal trauma, resuscitative endovascular balloon occlusion of the aorta (REBOA), and ResQFoam. The deductive thematic analysis aimed to answer three research questions. Research question 1 addressed the effectiveness of REBOA, damage control resuscitation, and damage control surgery in managing NCAH in austere environments. No effectiveness studies were found on this topic. Research question 2 identified three knowledge gaps in NCAH management in austere environments. The analysis identified early hemorrhage control, prehospital provider decision-making ability, and REBOA implementation as knowledge gaps in NCAH. Research question 3 identified five innovations that may affect the management of NCAH in the future: transport of patients, advanced resuscitative care, expert consultation, REBOA implementation, and self-expanding foam implementation. The inductive thematic analysis resulted in four recurrent themes from the literature: prehospital care, decision-making, hemorrhage control, and mortality in NCAH. During the stakeholders' consultation, the results of the deductive and inductive thematic analyses were reviewed and agreed on by the stakeholders. Special emphasis and discussion were given to prehospital management, expert opinions in the prehospital environment, decision-making in the prehospital environment, transport and resuscitation in the prehospital setting, REBOA, alternative discussion for research, and research gaps., Conclusion: NCAH is still a significant cause of preventable death in both military and civilian austere environments, even with ongoing research and interventions aimed at extending survival in such conditions. This scoping review has identified several potential concepts that could reduce the mortality associated with a preventable cause of death due to hemorrhage in austere environments., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: To evaluate refractive and topographic results of the association of intrastromal corneal ring segments (ICRS) with photorefractive keratectomy (PRK) for the correction of high (>6.0 diopters [D]) postkeratoplasty astigmatism (PKA)., Setting: University of São Paulo, São Paulo, Brazil., Design: Prospective interventional study., Methods: Postpenetrating keratoplasty patients, intolerant to contact lens fitting, and with corneal astigmatism higher than 6.0 D were treated by the combination of ICRS and PRK from January 2017 to June 2019. First, patients underwent femtosecond laser-assisted ICRS implantation to reduce and regularize corneal astigmatism, and 3 months later, submitted to PRK for the residual astigmatism. Outcomes were obtained 12 months after PRK., Results: The study comprised 30 eyes of 29 patients. Mean uncorrected distance visual acuity (logMAR) changed from 1.16 ± 0.37 in the preoperative to 0.69 ± 0.40 after ICRS ( P < .0001) and to 0.34 ± 0.29 12 months after PRK ( P < .0001). Mean spherical equivalent decreased from -5.19 ± 4.81 D in the preoperative to -3.38 ± 4.51 D after ICRS ( P < .0001) and to -2.30 ± 2.84 D after PRK ( P = .132). Mean topographic astigmatism decreased from 7.88 ± 2.13 D in the preoperative to 5.47 ± 2.29 D after ICRS ( P < .0001) and to 4.12 ± 2.93 D after PRK ( P = .003). Mean refractive astigmatism decreased from 7.10 ± 1.13 D in the preoperative to 4.61 ± 1.61 D after ICRS ( P < .0001) and to 2.58 ± 1.49 D after PRK ( P < .0001). After PRK, the mean correction index (CI) for corneal astigmatism was 0.77 ± 0.36. The ICRS/PRK combination resulted in a higher CI than ICRS only, both for corneal and refractive astigmatism. 2 eyes (8%) presented clinically significant opacification. Other complications were endothelial rejection (n = 1, 4%), infectious keratitis (n = 1, 4%), and ICRS extrusion after corneal melting (n = 1, 4%)., Conclusions: The association of ICRS and PRK was effective for treating high PKA. This strategy improved visual acuity, spherical equivalent, topographic and refractive astigmatism and resulted in a high CI. Safety questions remain open and must be balanced against benefits., (Copyright © 2022 Published by Wolters Kluwer on behalf of ASCRS and ESCRS.)

Objective: To analyze the patent registrations registered in the nursing area from 2004 to 2019, relating to the beginning of the Professional Masters in Nursing., Method: Electronic documentary research carried out at the bases of the National Institute of Industrial Property and Lattes Platform. After data collection, requests, filing date, patent titles, international patent classification and areas of expertise of depositor / inventors were analyzed., Results: The study points out that there was no significant correlation between the number of registered patents with the prelude to the professional master's degree in nursing, corroborating with previous studies regarding incipience., Final Considerations: although two of the initial objectives of professional master's programs are creation and technological innovation, it is analyzed that the granted institutions need to simplify and equip researchers with regard to the patent registration process, so that we can take off in this technological universe.

Objective: To reflect on the key concepts of the book The Structure of Scientific Revolutions and its applicability in professional master's in nursing., Method: This is a theoretical-reflective study that uses the philosophical and epistemological conceptions of the philosopher Thomas Samuel Kuhn to consider its applicability on the paradigm shift of stricto sensu graduate courses in nursing. The main concepts of Kuhn were used as support: paradigm, anomaly, scientific community and scientific revolution., Results: The propositions of this philosopher are applied to and support the theoretical reflection on professional master's programs, contributing to clarify what would be a paradigmatic visionary perspective in stricto sensu master's models in nursing., Conclusion: From Kuhn's propositions it was possible to conclude that professional master's programs in nursing can break away from the dominant paradigm, strengthening a scientific revolution within the academia.

A radiological dispersive device (RDD) spreads radioactive material, complicates the treatment of physical injuries, raises cancer risk, and induces disproportionate fear. Simulating such an event enables more effective and efficient utilization of the triage and treatment resources of staff, facilities, and space. Fast simulation can give detail on events in progress or future events. The resources for triage and treatment of contaminated trauma victims can differ for pure exposure individuals, while discouraging the "worried well" from presenting in the crisis phase by media announcement would relieve pressure on hospital facilities. The proposed methodology integrates capabilities from different platforms in a convergent way composed of three phases: (a) scenario simulation, (b) data generation, and (c) risk assessment for triage focused on follow-up epidemiological assessment. Simulations typically indicate that most of the affected population does not require immediate medical assistance. Medical triage for the few severely injured and the radiological triage to diminish the contamination with radioactivity will always be the priority. For this study, however, higher priorities should be given to individuals from radiological "warm" and "hot" zones as required by risk criteria. The proposed methodology could thus help to (a) filter and reduce the number of individuals to be attended, (b) optimize the prioritization of medical care,, ((c) reduce or prepare for future costs, (d) effectively locate the operational triage site to avoid possible contamination on the main facility, and (e) provide the scientific data needed to develop an adequate approach to risk and its proper communication.)

Objective: To summarize the production of the Professional Master's Program in Nursing Care Management of the Federal University of Santa Catarina, between 2013 and 2016., Method: electronic documental research. After data collection, we analyzed the numbers of defenses in relation to what was predicted by the respective public notices; as well as sex, training time and professional area of the authors; scenario, context and research line; general objective, analysis support model, methodological approach, instruments/techniques of data collection, and technique of analysis; and, finally, technological productions., Results: 57 dissertations were found and subjected to analysis. The highest number of defenses took place in 2016, in the public scenario, in a care context, with a qualitative approach and having assistance protocols as a final product., Conclusion: Although the country has weaknesses in its educational system, results of the post-graduate level stand out through the technological productions of professional master's studies in nursing.

In this study, interactions between polypeptide-leucine (0.2% w/w) and gum arabic (0.03, 0.06, 0.09, 0.12, and 0.15% w/w) were examined at concentrations of NaCl (0, 0.01, 0.25, 0.3, 0.5mol/l) and at different pH values (from 1.0 to 12.0). Formation of insoluble complex coacervates was highest at pH 4.0. At pH 2.0, which is the pKa of the gum Arabic, the dissociation of precipitate occurred. The pHØ2 positively shifted with the addition of higher concentrations of salt. Samples containing 0.2% PL and 0.03% GA and no salt had higher turbidity and increased formation of precipitates showing greater turbidity and particle sizes. The Fourier transform infrared spectroscopy confirms the complex coacervate formation of leucine and gum arabic, and rheological measurements suggest the elastic behavior of 0.2% PL and 0.03% GA complex. Overall, the study suggests that complex coacervates of PLs could be one feasible ways of incorporating amino acids in food products., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objectives: Assess the worthiness of arthroscopy in investigating and treating knee pain after arthroplasty unexplained by clinical and subsidiary examinations., Methods: Among 402 patients submitted to total or unicompartimental arthroplasty between September 2001 and April 2007 at a public university hospital, 17 presented with pain on prosthetic articulation, without clear diagnosis by clinical, X-ray, laboratory, scintiscan, or nuclear magnetic resonance tests. All patients were submitted to arthroscopy and symptoms were assessed by using the Lysholm scale, comparing pre-and post-arthroscopy periods. Peroperative findings have been recorded., Results: The procedure was effective for pain relief in 14 of 17 patients (82.35%). The median for Lysholm scale climbed from 36 points before arthroscopy to 94 points after the procedure (p < 0.001). Most of the patients (12) were arthroscopically diagnosed with fibrosis known as "cyclop"; on the remaining five patients, anterior synovitis was found. All patients were treated by resection., Conclusions: Knee arthroscopy after arthroplasty in patients presenting unclear persistent pain shows localized arthrofibrosis ("cyclops") or synovitis, which can be treated by using the same procedure, resulting in pain relief.

Background: Several substances have been researched to act as vehicles associated with calcium hydroxide. The specific type of vehicle is directly related to the effectiveness of the ionic dissociation, antimicrobial action, and biocompatibility of this medication., Aim: To make a histological evaluation of the biocompatibility of calcium hydroxide associated with a new vehicle (triethanolamine), compared with polyethylene glycol, saline solution, and olive oil., Materials and Methods: Fifty mice of guinea pig species were randomly divided into five groups (n = 10) according to each vehicle used--Group 1: calcium hydroxide, Group 2: triethanolamine, Group 3: polyethylene glycol, Group 4: saline solution, and Group 5: olive oil--and further divided into subgroups according to the two analysis periods--(a) 30 and (b) 90 days. Teflon carriers filled with the evaluated substances were placed in standardized bone cavities in the anterior mandible region. The animals were euthanized to perform a histological analysis after the time periods analyzed., Results: In 30 days, specimens from Groups 1, 3, and 5 showed a very pronounced inflammatory response. Specimens from Group 2 showed an inflammatory reaction ranging from mild to severe, with rapid resorption of the material and progressive advancement of osteoid tissue into the teflon carriers. Specimens from Group 4 showed a moderate inflammatory reaction. In 90 days, specimens from Group 1 showed a very pronounced fibrous replacement. In regard to Group 2 specimens, the tested material was solubilized and replaced by newly formed bone tissue. For Groups 3 and 5 specimens, the inflammatory reaction went from acute to moderate. In relation to Group 4 specimens, an organized bone formation process was observed., Conclusions: Specimens from Group 2 showed higher biocompatibility, especially as compared with the specimens from Groups 3 and 5.

In spite of extensive research, immunologic control mechanisms against Porcine Reproductive and Respiratory Syndrome virus (PRRSv) remain poorly understood. Cytokine responses have been exhaustively studied in nursery pigs and show contradictory results. Since no detailed reports on cytokine responses to PRRSv in pregnant females exist, the objectives of this study were to compare host cytokine responses between PRRSv-infected and non-infected pregnant gilts, and to investigate relationships between cytokine levels in infected gilts and viral load or fetal mortality rate. Serum samples and supernatants of peripheral blood mononuclear cells (PBMC) either stimulated with PRRSv or phorbol myristate acetate/Ionomycin (PMA/Iono) were analyzed for cytokines/chemokines: interleukins (IL) 1-beta (IL1β), IL4, IL8, IL10, IL12, chemokine ligand 2 (CCL2), interferon alpha (IFNα) and interferon gamma (IFNγ). Three cytokines (IFNα, CCL2, IFNγ) in gilt serum differed significantly in inoculated versus control gilts over time. In supernatants of PRRSv stimulated PBMC from PRRSv-infected gilts, levels of IFNα were significantly decreased, while IL8 secretion was significantly increased. PRRSv infection altered the secretion of all measured cytokines, with the exception of IFNα, from PBMC after mitogen stimulation, indicating a possible immunomodulatory effect of PRRSv. IFNα, CCL2, and IFNγ in serum, and IFNγ in supernatants of PMA/Iono stimulated PBMC were significantly associated with viral load in tissues, serum or both. However, only IFNα in supernatants of PRRSv stimulated PBMC was significantly associated with fetal mortality rate. We conclude that of the eight cytokines tested in this study IFNα was the best indicator of viral load and severity of reproductive PRRSv infection.

Litter size (LS) in sheep is determined mainly by ovulation rate (OR). Several polymorphisms have been identified in the growth differentiation factor 9 (GDF9) gene that result in an increase in OR and prolificacy of sheep. Screening the databank of the Brazilian Sheep Breeders Association for triplet delivery, we identified flocks of prolific Ile de France ewes. After resequencing of GDF9, a point mutation (c.943C>T) was identified, resulting in a non-conservative amino acid change (p.Arg315Cys) in the cleavage site of the propeptide. This new allele was called Vacaria (FecG(v) ). A flock of half-sib ewes was evaluated for OR in the first three breeding seasons, and Vacaria heterozygotes had higher OR (P < 0.001), averaging 2.1 ± 0.1 when compared to 1.2 ± 0.1 in wild-type ewes. The OR was also influenced by age, increasing in the second and third breeding seasons (P < 0.001). In flocks segregating this allele, the LS was higher in mutant sheep (P < 0.001), averaging 1.61 ± 0.07 in heterozygotes and 1.29 ± 0.03 in wild-type ewes. Analysis of homozygote reproductive tract morphology revealed uterine and ovarian hypoplasia. Ovarian follicles continue to develop up to small antral stages, although with abnormal oocyte morphology and altered arrangement of granulosa cells. After the collapse of the oocyte in most follicles, the remaining cells formed clusters that persisted in the ovary. This SNP is useful to improve selection for dam prolificacy and also as a model to investigate GDF9 post-translation processing and the fate of the follicular cells that remain after the oocyte demise., (© 2014 Stichting International Foundation for Animal Genetics.)

The conversion of lignocellulose into fermentable sugars is considered a promising alternative for increasing ethanol production. Higher fermentation yield has been achieved through the process of simultaneous saccharification and fermentation (SSF). In this study, a comparison was performed between the yeast species Saccharomyces cerevisiae and Kluyveromyces marxianus for their potential use in SSF process. Three strains of S. cerevisiae were evaluated: two are widely used in the Brazilian ethanol industry (CAT-1 and PE-2), and one has been isolated based on its capacity to grow and ferment at 42 °C (LBM-1). In addition, we used thermotolerant strains of K. marxianus. Two strains were obtained from biological collections, ATCC 8554 and CCT 4086, and one strain was isolated based on its fermentative capacity (UFV-3). SSF experiments revealed that S. cerevisiae industrial strains (CAT-1 and PE-2) have the potential to produce cellulosic ethanol once ethanol had presented yields similar to yields from thermotolerant strains. The industrial strains are more tolerant to ethanol and had already been adapted to industrial conditions. Moreover, the study shows that although the K. marxianus strains have fermentative capacities similar to strains of S. cerevisiae, they have low tolerance to ethanol. This characteristic is an important target for enhancing the performance of this yeast in ethanol production.

An extracellular β-glucanase secreted by Kluyveromyces marxianus was identified for the first time. The optimal conditions for the production of this enzyme were evaluated by response surface methodology. The optimal conditions to produce β-glucanase were a glucose concentration of 4% (w/v), a pH of 5.5, and an incubation temperature of 35 °C. Response surface methodology was also used to determine the pH and temperature required for the optimal enzymatic activity. The highest enzyme activity was obtained at a pH of 5.5 and a temperature of 55 °C. Furthermore, the enzyme was partially purified and sequenced, and its specificity for different substrates was evaluated. The results suggest that the enzyme is an endo-β-1,3(4)-glucanase. After optimizing the conditions for β-glucanase production, the culture supernatant was found to be effective in digesting the cell wall of the yeast Saccharomyces cerevisiae, showing the great potential of β-glucanase in the biotechnological production of soluble β-glucan.

The aim of this study was to evaluate the bond strength to the dentin of an adhesive material used for root reinforcement light activated with different sources. Roots were divided into 4 groups (n=15) according to the light source used to activate the resin reinforcement: GI, non-weakened roots (control); GII, halogen light (H) 600 mW/cm²; GIII, LED 800 mW/cm² and GIV, LED 1500 mW/cm². The reinforcement was done with adhesive, composite resin and fiberglass posts. After 24 h, the specimens were sectioned and the first slice of each post region was used in the push out test in a universal testing machine with a crosshead speed of 0.5 mm/min. Failure modes of the debonded specimens were examined. Data (MPa) were analyzed by ANOVA and Holm-Sidak test (α=0.05). The second slice from each region was analyzed by scanning electron microscopy (SEM). LED-1500 (4.69 ± 1.74) provided bond strength similar to the control group (5.05 ± 2.63) and statistically different from H-600 (1.96 ± 0.94) and LED-800 (2.75 ± 1.90), which were similar to each other (p<0.05). Cervical (4.16 ± 2.32) and middle (4.43 ± 2.32) regions showed higher bond strength than the apical (2.25 ± 1.50) (p<0.05). There was a prevalence of adhesive failures in H-600 and LED-800 and cohesive failures in LED-1500. SEM showed the formation of long, numerous and fine tags. It was concluded that LED-1500 provided higher bond strength of resin reinforcement to the dentin.

Bead-based multiplex assays (BBMAs) are applicable for high throughput, simultaneous detection of multiple analytes in solution (from several to 50-500 analytes within a single, small sample volume). Currently, few assays are commercially available for veterinary applications, but they are available to identify and measure various cytokines, growth factors and their receptors, inflammatory proteins, kinases and inhibitors, neurobiology proteins, and pathogens and antibodies in human beings, nonhuman primates, and rodent species. In veterinary medicine, various nucleic acid and protein-coupled beads can be used in, or for the development of, antigen and antibody BBMAs, with the advantage that more data can be collected using approximately the same amount of labor as used for other antigen and antibody assays. Veterinary-related BBMAs could be used for detection of pathogens, genotyping, measurement of hormone levels, and in disease surveillance and vaccine assessment. It will be important to evaluate whether BBMAs are "fit for purpose," how costs and efficiencies compare between assays, which assays are published or commercially available for specific veterinary applications, and what procedures are involved in the development of the assays. It is expected that many veterinary-related BBMAs will be published and/or become commercially available in the next few years. The current review summarizes the BBMA technology and some of the currently available BBMAs developed for veterinary settings. Some of the human diagnostic BBMAs are also described, providing an example of possible templates for future development of new veterinary-related BBMAs.

Objective: To evaluate whether treatment with L-arginine influences the healing of skin flaps in rats exposed to nicotine., Methods: 40 male Wistar rats weighing 142.4 ± 10.1 g were separated into four groups: GC: treatment with 7.4 pH phosphate buffer, submitted to skin flap and observation for ten days; GN: exposure to nicotine for four weeks, submitted to skin flap and observation for ten days; GA: treatment with 7.4 pH phosphate buffer for four weeks, submitted to skin flap and arginine treatment for ten days; GAN: exposure to nicotine for four weeks, submitted to skin flap and treatment with arginine for ten days. We evaluated: areas of necrosis, re-epithelialization, inflammatory reaction and formation of granulation tissue by HE stain; the total area of deposition and differentiation of collagens I and III by histometry with picrosirius staining; and the scar vascular density by immunohistochemical staining with monoclonal anti-CD34 antibodies., Results: The percentages of necrotic areas in GN and GNA were higher (p <0.001) than in GC and GA. In histological scores, collagen deposition, and the percentage of type I collagen, GA and GC were similar to each other (p> 0.05), but higher (p <0.001) than GA and GNA; as for vascular densities, they were lower in GN and GAN (p <0.001) than in GC and GA., Conclusion: Exposure to nicotine inhibited the effects of arginine and in unexposed mice there was induction of angiogenesis and improvement in the total collagen deposition in the skin flaps.

Purpose: Inadequate iodine intake may result in iodine deficiency disorders (IDD). Thus, for more than 50 years, policies for the regulation of salt fortification with iodine have existed in Brazil. In 2003, a study on 6-14-year-old schoolchildren from regions of the state of São Paulo showed a median urinary iodine concentration of 360 μg/L. The objective of the present study was to assess the iodine nutrition status among schoolchildren., Methods: The study was conducted on 828 schoolchildren aged 4-13 years from eight schools in the interior of the state of São Paulo. A casual urine sample was collected from each volunteer for iodine determination by the adapted method of Sandell-Kalthoff., Results: Only 1.9% (n = 16) of the children evaluated had low values of urinary iodine (<100 μg/L), while 24.6% had urinary iodine excretion values between 200 and 300 μg/L, and 67.1% had values above >300 μg/L., Conclusions: The results show that the iodine nutritional status of the schoolchildren studied is characterized by a high urinary iodine excretion, which might reveal an increase in iodine consumption by this population.

Ethanol can be produced from cellulosic biomass in a process known as simultaneous saccharification and fermentation (SSF). The presence of yeast together with the cellulolytic enzyme complex reduces the accumulation of sugars within the reactor, increasing the ethanol yield and saccharification rate. This paper reports the isolation of Saccharomyces cerevisiae LBM-1, a strain capable of growth at 42 °C. In addition, S. cerevisiae LBM-1 and Kluyveromyces marxianus UFV-3 were able to ferment sugar cane bagasse in SSF processes at 37 and 42 °C. Higher ethanol yields were observed when fermentation was initiated after presaccharification at 50°C than at 37 or 42° C. Furthermore, the volumetric productivity of fermentation increased with presaccharification time, from 0.43 g/L/h at 0 h to 1.79 g/L/h after 72 h of presaccharification. The results suggest that the use of thermotolerant yeasts and a presaccharification stage are key to increasing yields in this process., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Coutarea hexandra is a species commonly known in Brazil as quina, and its bark is used in folk medicine. In this study, we assess the mutagenic and DNA-damaging effects of ethanol extracts from C. hexandra stem bark (SCH) and leaves (LCH) by employing the Ames test on the TA98 and TA100 strains of Salmonella typhimurium in addition to a plasmid treatment test. Furthermore, we performed a phytochemical analysis by TLC and HPLC, a quantification of the phenolic constituents and an assessment of the antioxidative activity. SCH and LCH showed mutagenic action in the Ames test for TA98 strains after metabolic activation. LCH also showed mutagenicity for the TA100 strain after metabolic activation. The findings from the plasmid treatment test did not indicate any DNA-damaging activity for either of the extracts with the tested dosages. SCH showed greater flavonoid content and greater antioxidative potential in relation to LCH. This study suggests that caution is advisable in the use of this plant. However, in vivo studies should be conducted to confirm these data., (Copyright © 2011 Elsevier B.V. All rights reserved.)

One of the hallmarks of polycystic ovary syndrome (PCOS) is increased ovarian androgen secretion that contributes to the ovarian, hormonal, and metabolic features of this condition. Thecal cells from women with PCOS have an enhanced capacity for androgen synthesis. To investigate whether this propensity is a potential cause, rather than a consequence, of PCOS, we used an ovine prenatal androgenization model of PCOS and assessed ewes at 11 months of age. Pregnant Scottish Greyface ewes were administered 100 mg testosterone propionate (TP) or vehicle control twice weekly from d 62 to 102 of gestation, and female offspring (TP = 9, control = 5) were studied. Prenatal TP exposure did not alter ovarian morphology or cyclicity, or plasma androgen, estrogen, and gonadotropin concentrations, at this stage. However, follicle function was reprogrammed in vivo with increased proportions of estrogenic follicles (P < 0.05) in the TP-exposed cohort. Furthermore, in vitro the thecal cells of follicles (>4 mm) secreted more LH-stimulated androstenedione after prenatal androgenization (P < 0.05), associated with increased basal expression of thecal StAR (P < 0.01), CYP11A (P < 0.05), HSD3B1 (P < 0.01), CYP17 (P < 0.05), and LHR (P < 0.05). This provides the first evidence of increased thecal androgenic capacity in the absence of a PCOS phenotype, suggesting a thecal defect induced during fetal life.

Objective: To determine whether tacrolimus administered to rats, in the presence of pancreatitis induced by L-Arginine, interferes with the serum levels of amylase and glucose and the histological pattern of the pancreatic parenchyma., Methods: Forty Wistar rats were divided into four groups with 10 rats each: control group (C), tacrolimus group (T), pancreatitis group (P) and pancreatitis-tacrolimus group (PT). We evaluated serum levels of amylase, glucose, and tacrolimus and made histological assessments of the pancreas. Induction of pancreatitis was made by inoculation of L-Arginine at a dose of 500 mg/100g body weight intraperitoneally, and tacrolimus treatment at a dose of 1ìg/kg subcutaneously for four days., Results: Serum amylase was higher (p = 0.0000) in groups PT, P and T than in the control group. The PT group mean was higher (p = 0.0009) than in the T group, but did not differ (p = 0.6802) from the average of the P group. There was no difference between groups P and T (p = 0.2568). Neither in mean blood glucose between the groups (p = 0.4920); serum levels of tacrolimus were similar in PT and T groups (p = 0.7112). There were no histological changes in groups T and C and no hemorrhage in the pancreas of rats in groups P and PT. In group P, there was no edema in 30%, mild edema in 20% and in 50%, moderate; as for inflammatory infiltration, it was moderate in 80% and absent in 20%, and atrophy of the parenchyma was moderate in 60% and severe in 40%. In the PT group, there was edema, inflammatory infiltration or atrophy in the pancreas in all rats., Conclusion: Treatment with Tacrolimus induced an increase in serum amylase in normal mice, but did not affect blood glucose or the histological pattern of the pancreatic parenchyma. In the presence of pancreatitis induced by L-Arginine tacrolimus induced edema, inflammatory infiltration and more severe atrophy in the pancreatic parenchyma.

Brazilian Santa Inês (SI) sheep are very well-adapted to the tropical conditions of Brazil and are an important source of animal protein. A high rate of twin births was reported in some SI flocks. Growth and Differentiation Factor 9 (GDF9) and Bone Morphogenetic Protein 15 (BMP15) are the first two genes expressed by the oocyte to be associated with an increased ovulation rate in sheep. All GDF9 and BMP15 variants characterized, until now, present the same phenotype: the heterozygote ewes have an increased ovulation rate and the mutated homozygotes are sterile. In this study, we have found a new allele of GDF9, named FecG(E) (Embrapa), which leads to a substitution of a phenylalanine with a cysteine in a conservative position of the mature peptide. Homozygote ewes presenting the FecG(E) allele have shown an increase in their ovulation rate (82%) and prolificacy (58%). This new phenotype can be very useful in better understanding the genetic control of follicular development; the mechanisms involved in the control of ovulation rate in mammals; and for the improvement of sheep production., (© 2010 The Authors, Animal Genetics © 2010 Stichting International Foundation for Animal Genetics.)

The halo 6-fatty acid esters of L-ascorbic acid 3a, 3b and 6-fatty acid esters of L-ascorbic acid 5a-g were achieved from L-ascorbic acid 1. Compounds 3a, 3b and 5a-g were evaluated for anti-oxidant, anti-lipid peroxidation, and secretory phospholipase A(2) (sPLA(2)) inhibition in vitro, and sPLA(2) induced mouse paw edema. All the derivatives retained their anti-oxidant property compared to ascorbic acid at 6 × 10(-4)M and are good inhibitors of lipid peroxidation at 1 mg ml(-1) as evaluated by 2, 2-Diphenyl-1-picrylhydrazyl radical and thio-barbituric acid methods, respectively. Compounds 5e and 5f significantly inhibited purified group I sPLA(2) from Naja naja and group II sPLA(2) from Vipera russelli, human synovial fluid and human pleural fluid with IC(50) value ranging from 64 ± 1.95 to 82 ± 1.3 and 48 ± 2.27 to 61 ± 2.23 μM, respectively. The compounds 5e and 5f also showed varying degree of potency in neutralizing indirect hemolytic activity of sPLA(2) at 50 μM concentration, and sPLA(2) induced mouse paw edema at the dose 3 mg/kg. Further docking studies also confirmed that compounds 5e and 5f have maximum interaction with increasing negative energy value. Single molecule possessing both anti-oxidant and anti-inflammatory activities is of great therapeutic significance in inflammatory disorders.

The objective of the present study was to evaluate the urinary excretion of iodine and relate it to the amount present in salt for human consumption. The study involved 145 children from two schools: a rural one and an urban one. We performed anthropometric measurements and collected a urine sample and a kitchen salt sample from each child. In the rural school, 3.38% of children had iodine deficiency. However, most of the values of urinary iodine were above 300 microg/L (62.03%) and 59.49% of the kitchen salt samples contained 20 to 60 mg iodine per kilo of salt. In the urban school, 3.03% of the children had urinary iodine excretion of less than 100 microg/L and 90.91% of the children had urinary iodine values exceeding 300 microg/L. Ofthis total, 84.85% of the kitchen salt samples contained 20 to 60 mg iodine per kilo of salt. Iodine deficiency is controlled in this population, with the current reality showing a high prevalence of excess urinary iodine.

Two experiments were carried out on ewes with ovarian autotransplants to estimate the ovarian uptake of glucose and production of lactate. The first was carried out in the luteal phase of the oestrous cycle. Samples of carotid arterial, ovarian venous and jugular venous blood were collected simultaneously for glucose analysis. The arterial concentration of glucose (58.0 +/- 5.0mg/dL; Mean+/-SEM) was significantly higher than the ovarian venous concentration (42.3+/-2.4 mg/dL; P<0.001). Next, a second more complete experiment was carried out in the luteal and follicular phases of the oestrous cycle. The oestrous cycle was synchronised and samples of carotid arterial, ovarian venous and jugular venous blood were collected simultaneously for glucose and lactate analysis. There were significant positive arterio-venous differences in the concentration of glucose in the luteal (5.6+/-1.2mg/dL, mean+/-SEM; P=0.001), early (3.1+/-0.82 mg/d; P=0.003) and late follicular (6.4+/-1.3mg/dL; P=0.001) phases of the oestrous cycle. There was a significant negative arterio-ovarian venous difference in the concentration of lactate in only the luteal phase (-2.2+/-0.96 mg/dL; P=0.043). The results show significant removal of glucose from the arterial circulation during its passage through the ovary in the luteal, early follicular and late follicular phases of the oestrous cycle. Furthermore, there was lactate production in the luteal phase but not in the follicular phase suggesting that in the luteal phase of the oestrous cycle, ovarian metabolism can be anaerobic., ((c) 2010. Published by Elsevier Inc.)

Background: Periapical lesions are a host response that involves immune reaction to prevent dissemination of bacteria from an infected root canal. The purpose of this study was to evaluate the levels of nitric oxide (NO), IL-4, TGF-beta, tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) in chronic periapical lesions and to determine their possible association with clinical and radiographic parameters., Methods: Seventeen human radicular cysts and 30 periapical granulomas were used in this study. Cytokines and NO were assessed by enzyme-linked immunosorbent assay and by the Griess reaction respectively confirmed by immunohistochemical., Results: TNF-alpha and IFN-gamma were detected in 10% of granulomas and in 41.2% and 70% of radicular cysts. IL-4 was reactive in 24% of cysts, and TGF-beta was positive in all samples. Patients with tenderness showed significantly higher levels of IFN-gamma and IL-4 (P < 0.05). Swelling was associated with high levels of TNF-alpha, IFN-gamma, and IL-4 (P < 0.05). Lesions presenting bone resorption were associated with high levels of NO (P < 0.05)., Conclusions: Periapical granulomas display a regulatory environment characterized by high TGF-beta and low inflammatory cytokine levels, while radicular cysts has mist Th1 and Th2 inflammatory reaction with the presence of IFN-gamma, TNF-alpha, and IL-4.

This study was designed to assess possible associations between biomarkers of mercury (Hg) exposure and oxidative stress in fish-eating Amazonian communities. Clinical samples were obtained from riparians living in the Brazilian Amazon. Biomarkers of oxidative stress (glutathione - GSH, glutathione peroxidase - GSH-Px, catalase - CAT, activity and reactivation index of delta-aminolevulinate dehydratase - ALA-D (R%) were determined in blood. Total Hg was measured in whole blood (B-Hg), plasma (P-Hg) and hair (H-Hg). Association between biomarkers of Hg exposure and oxidative stress were examined using multiple regression models, including age, gender, alcohol consumption, smoking status, fish consumption and then stratified for gender. Significant inverse relations were observed between GSH-Px, GSH, CAT, ALA-D activity and B-Hg or H-Hg (p<0.05). ALA-D reactivation index was positively related to B-Hg (p<0.0001). P-Hg was directly related to ALA-D reactivation index and inversely associated with GSH-Px, GSH, and ALA-D activity (p<0.05). When stratified for gender, women showed significant inverse associations between all biomarkers of Hg exposure and CAT (p<0.05) or GSH (p<0.05), while for men only P-Hg showed a significant inverse relation with GSH (p<0.001). Our results clearly demonstrated an association between Hg exposure and oxidative stress. Moreover, for B-Hg, P-Hg and H-Hg gender differences were present., (Copyright 2009 Elsevier B.V. All rights reserved.)

In this review, we have compiled the data on pharmacological activities associated with endogenous purine release related enzymes-nucleases (DNases, RNases, and phosphodiesterases). The results of studies on toxic effects of these enzymes, emphasizing the future directions in this field, are summarized. One of the major problems facing toxicologists is the identification and characterization of specific venom nucleases since they share similar substrate specificities and biochemical properties. In this review, we have attempted to clarify some of the discrepancies about these enzymes. Further, we have tried to correlate the existence of nuclease enzymes in relation to endogenous release of purines, a multitoxin, during snake envenomation, and we also discuss the possible actions of purines. We hope that this review will stimulate renewed interest among toxicologists to biologically characterize these enzymes and elucidate their role in envenomation.

The purpose of this study was to compare the accuracy of Root ZX II to locate the apical constriction with the display meter set at "0.5" and "1'"reading. Seventy single-rooted teeth were soaked in an alginate model and randomly distributed in 2 groups (n = 35). Measurements were taken following canal irrigation with 1% NaOCl. The length was established using a #20 K-file attached to the holder when the display indicator reached the marks "0.5" (group I) or "1" (group II), after the meter read "Apex." Then, the file was fixed in position and the teeth removed from the alginate. The apical portion of the root was shaved until the tip of the file could be seen, the distance to the apical constriction verified by means of a stereomicroscope and the measurements compared. Statistical analysis was performed by using Student t test with the null hypothesis set as 5%. The mean positions of the file tip relative to the apical constriction were -0.23 +/- 0.39 mm and -0.42 +/- 0.45 for groups I and II, respectively, with no statistical difference (P > .05). The accuracy was 90.5% and 83.78% for the Root ZX II "0.5" and "1" readings, respectively. It was concluded that the meter reading "1" of Root ZX II reduced the risk of working length overestimation.

Objective: To evaluate the individual functionality of gonadotropin-stimulated preovulatory follicles, for understanding embryo failure in assisted reproductive technique cycles, in a sheep model., Design: Observational, model study., Setting: Public research unit., Animal(s): Fifteen adult Manchega ewes., Intervention(s): Synchronization of the estrous cycle with intravaginal progestagens and ovarian stimulation with FSH; evaluation of reproductive activity, plasma sampling, ovarian ultrasonography, and ovariectomies., Main Outcome Measure(s): Determination of estrus behavior, plasma and intrafollicular concentrations of E(2) and inhibin A, number and size of ovarian follicles, and developmental competence of oocytes., Result(s): These results support the usefulness of serial measurements of plasma inhibin A for assessment of follicular growth during the FSH treatment, rather than of E(2) assays commonly used. Functionality of FSH-stimulated preovulatory follicles is clearly disturbed, as confirmed by a negative correlation between follicular size and intrafollicular concentrations of inhibin A and E(2) in preovulatory follicles after individual dissection; moreover, the ability of their oocytes to resume meiosis was diminished., Conclusion(s): Functionality of follicles in controlled ovarian stimulation (COS), and developmental competence of their oocytes, is disturbed by the high doses of gonadotropin supplied and finally determined by follicular sizes at starting FSH treatment.

Background: Renal abnormalities in leprosy have been largely described in medical literature, but there are few studies evaluating renal function in these patients., Methods: This is a cross-sectional study in 59 consecutive paucibacillary (PB) and multibacillary (MB) leprosy patients. Glomerular filtration rate (GFR) was estimated by simplified-MDRD formula. Microalbuminuria was determined by 24 h urine collection. Urinary acidification capacity was measured after water deprivation and acid-loading with CaCl(2). Urinary concentration capacity was evaluated after desmopressin acetate administration, using the urinary to plasma osmolality (U/P(osm)) ratio. All parameters except microalbuminuria were measured in a control group of 18 healthy volunteers., Results: Age and gender were similar between leprosy (MB or PB) and control groups. GFR 1.2 mg/dl was observed in 17.9% of MB patients and in none of the controls (P=0.020). A negative correlation was observed between GFR and time of treatment (r= -0.339; P=0.002). Age and time of treatment were independent risk factors for GFR

Objective: To evaluate the subcutaneous biocompatibility of 2 root canal sealers., Study Design: The subcutaneous implant technique recommended by the Fédération Dentaire International (FDI) was used to test Endométhasone and EndoREZ root canal sealers. These materials were placed in Teflon tubes, 1 mm in diameter and 10 mm in length, and implanted into 2 pockets created in the back of 40 Calomys callosus rodents, 20 for each material. Tissue biopsies were collected and histologically examined 15, 30, 60, and 90 days after the implantation procedure. The overall level of the inflammatory tissue response was graded as none, slight, moderate, or severe on the sealer-connective tissue interface at the opening ends of the tubes. The connective tissue response along the lateral wall outside of each tube served as a negative control., Results: The tissue reaction to the Endométhasone diminished with time. The EndoREZ sealer was highly toxic during all experimental periods., Conclusion: Endométhasone root canal sealer presented biocompatibility within the analyzed periods, whereas EndoREZ showed no biocompatible behavior and caused late hypersensitive reaction.

Objective: To determine the effect of single long-acting doses of GnRH antagonists on reproductive function in a sheep model., Design: Observational, model study., Setting: University-affiliated research unit., Animal(s): Nine intact mature Merino sheep in experiment 1 and 12 mature Merino-crossed ewes with the ovary autotransplanted to the neck in experiment 2., Intervention(s): Synchronization of estrous cycle either with intravaginal progestins or prostaglandin F2alpha analogues and treatment with a single dose of GnRH antagonist; evaluation of reproductive activity, plasma sampling, and ovarian ultrasonography., Main Outcome Measure(s): Determination of estrus behavior; plasma concentrations of P, FSH, LH, and inhibin A; and number and size of ovarian follicles., Result(s): In both experiments, the concentrations of FSH and LH were suppressed when compared with those in control ewes. In experiment 1, the ovulatory cycles were suppressed for > or = 55 days in treated sheep. In experiment 2, there were no follicles sized > or = 5 mm in treated ewes for 50 days., Conclusion(s): The suppression of the development of large follicles for > or = 30 days after a single injection of a long-acting GnRH antagonist provides a novel convenient method of pretreatment before COS.

The FecB (Booroola) mutation, which leads to increased ovulation rates and multiple births in sheep, is now known to occur in the signaling domain of the bone morphogenic protein (BMP)-1B receptor. We examined the effect of the mutation on the responsiveness of granulosa (GC) and theca cells (TC) to BMPs and other local regulators using tissue from animals with (Fec(B/B)) and without (Fec(+/+)) the FecB mutation. Experiments examined the effect of BMP-2, -4, and -6 (0.005-50 ng/ml), and their interaction with IGF-I (0.1-10 ng/ml LR3 analog) and gonadotropins, on the proliferation and differentiation of GCs and TCs isolated from small (<2 mm) antral follicles and maintained in serum-free culture for up to 8 d. Dose-finding studies using ovaries from wild-type sheep obtained from the abbattoir showed no difference among the different BMPs in stimulating (P < 0.001) estradiol (E2) production by GCs cultured with FSH (10 ng/ml), but there was a clear interaction (P < 0.001) with IGF-I. BMPs had no effect on GC proliferation or the sensitivity of GCs to FSH. In contrast, higher doses of BMPs (5-50 ng/ml) inhibited LH-stimulated androstenedione production by TCs, whereas lower doses (0.005-0.05 ng/ml) stimulated TC proliferation (P < 0.01). Regardless of dose of IGF-I, at the end of culture (96-192 h) hormone production by GCs (E2, inhibin A) and TCs (androstenedione) was 4- to 5-fold greater (P < 0.001) by cells from Fec(B/B), compared with Fec(+/+) ewes exposed to the same dose of gonadotropin. In the presence of low concentrations of IGF-I (0.1 ng/ml), the maximum increase in the production of E2 and inhibin A by GCs from FF ewes in response to BMPs was observed at doses that were 3- to 10-fold lower (3-10 ng/ml) than ++ (30 ng/ml; P < 0.001). Low doses of BMPs stimulated proliferation of TCs from ++ (P < 0.01) but not FF ewes. Immunohistochemistry confirmed BMP-6 protein expression in the oocyte, granulosa, and thecal layers of antral follicles from both genotypes. These results confirm a major role for BMPs in controlling ovarian somatic cell function in sheep and provide evidence to support the hypothesis that the FecB mutation increases the BMP response of somatic cells when stimulated to differentiate by gonadotropins.

We have tested if the high number of unfertilized ova and degenerated embryos found in superovulated goats previously treated with GnRH antagonist can be related to a prolongation of gonadotrophin down-regulation and/or alterations in follicular function during the period of administration of the superovulatory treatment, around 4 days after the end of the antagonist treatment. A total of 15 does were treated with intravaginal progestagen sponges and daily injections of 0.5mg of the GnRH antagonist Antarelix for 6 days, while 5 does acted as controls receiving saline. During the antagonist treatment, the mean plasma LH concentration was lower in treated than control goats (0.5 +/- 0.2 versus 0.7 +/- 0.5 ng/ml, P < 0.0005 ); however, the FSH levels remained unaffected (0.8 +/- 0.4 versus 0.8 +/- 0.5 ng/ml). In this period, treated does also showed an increase in the number of small follicles with 2-3 mm in size ( 10.7 +/- 0.7 versus 8.4 +/- 0.6, P < 0.05), and a decrease in both the number of follicles > or =4 mm in size ( 5.0 +/- 0.3 versus 6.8 +/- 0.5, P < 0.005) and the secretion of inhibin A (120.9 +/- 10.7 versus 151.6 +/- 12.6 pg/ml, P < 0.05). After cessation of the antagonist treatment, there was an increase in LH levels in treated goats from the day after the last Antarelix injection (Day 1), so that LH levels were the same as controls on Day 3 (0.6 +/- 0.1 versus 0.6 +/- 0.2 ng/ml). However, there were even greater numbers of small follicles than during the period of antagonist injections (15.4 +/- 0.6 in treated versus 8.9 +/- 0.7 in control, P < 0.0005 ). Moreover, the number of > or =4 mm follicles and the secretion of inhibin A remained lower in treated goats (3.9 +/- 0.3 follicles and 84.4 +/- 7.0 pg/ml versus 5.4 +/- 0.5 follicles, P < 0.05 and 128.9 +/- 14.2 pg/ml, P < 0.05 ). These results indicate that pituitary secretion of gonadotrophins is restored shortly after the end of antagonist treatment, but activity of ovarian follicles is affected.

The objective was to study the endocrine activity in sheep with large ovarian follicles and the effects of dominant follicles on other follicles, looking for possible intraovarian differences. Induction of dominant follicles was achieved using controlled exogenous LH pulses every 90 min over 14 days in eight Scottish Blackface ewes. During this period, follicular development was assessed by daily transrectal ultrasonography and jugular venous blood samples were collected every 12 h for FSH, LH inhibin and oestradiol assay. The exogenous LH pulses caused the appearance of large follicles in all the ewes, which reached a maximum mean diameter of 7.2 +/- 0.5 mm on Day 5.5 +/- 2.6 after first detection. In the presence of a dominant follicle, no other follicle grew to a diameter larger than 4 mm and there was a decrease in the number of new growing follicles (P < 0.05) and in the number of smaller follicles (P < 0.01). This effect of dominance was mediated by changes in FSH concentration, since FSH level decreased (P < 0.05) as dominant follicles grew and the decrease in FSH levels was related to a decline in the number of remaining follicles (P < 0.05). However, the greatest decrease in the number of small follicles growing to larger sizes was observed in the ovary ipsilateral to the dominant follicle (P < 0.05). These data confirm that the presence of a large follicle depresses the recruitment and growth of other follicles by systemic factors and provide some evidence of local inhibitors blocking the final development of other putative large follicles.

It has been suggested that ewes carrying the Booroola gene (Fec(B)) consistently ovulate more follicles because they recruit more primordial follicles and/or have a lower rate of atresia. If the former is correct, the pool of follicles would be depleted sooner in Fec(B) animals. We have studied follicular dynamics and endocrine function during follicular and early luteal phases of the estrous cycle of older ewes with or without the fecundity gene and compared this data with data obtained 6 yr previously in the same animals. Older sheep carrying the Booroola gene maintained a significantly higher ovulation rate than noncarrier ewes [4.2 +/- 0.8 vs. 2.2 +/- 0.6 corpora lutea (CL), respectively; P < 0.05], and in keeping with data from young animals, both ovulatory follicles and CL (4.7 +/- 0.3 vs. 6.9 +/- 0.7 mm and 12.8 +/- 0.5 vs. 16.7 +/- 0.8 mm, respectively) were smaller than those of noncarrier ewes (P < 0.05). The interval from luteolysis to the onset of the LH surge increased with age in all the animals (from 52.0 +/- 8.0 to 67.0 +/- 7.5 h in gene carrier sheep and from 56.0 +/- 2.0 to 79.5 +/- 9.6 h in noncarrier sheep, P < 0.05). The concentration of estradiol and inhibin A in the early luteal phase was lower in older noncarrier ewes (P = 0.08 and P < 0.05, respectively), and the level of inhibin A was inversely related to the level of FSH in aged sheep of both genotypes (P < 0.0001). In contrast, the number of developing follicles in older ewes of both genotypes was similar to the number found in younger ewes, suggesting that increased ovulation rate in sheep carrying the Fec(B) mutation is related to a reduced rate of atresia.

The aim of this study was to determine, for goats, the effects of daily doses of GnRH antagonist on ovarian endocrine and follicular function. Ten does were given 45 mg FGA intravaginal sponges and then five were treated with daily injections of 0.5mg of the GnRH antagonist Teverelix for 11 days from 2 days after the day of sponge insertion, while five does acted as controls. Pituitary activity was monitored by measuring plasma FSH and LH daily from 2 days before the first GnRH injection to Day 12. Follicular activity was determined by ultrasonographic monitoring and by assessing plasma inhibin A levels during the same period. In treated does, the FSH levels decreased linearly (0.8 +/- 0.1 ng/ml to 0.5 +/- 0.1 ng/ml, P < 0.01) and remained lower than the mean concentration in control goats (0.8 +/- 0.1 ng/ml, P < 0.005). LH levels were also lower during the period of antagonist treatment (0.6 +/- 0.2 ng/ml versus 0.4 +/- 0.1 ng/ml, P < 0.0005). During GnRH antagonist treatment, there was a significant decrease in the number of large follicles (> or = 6 mm) from Day 3 of treatment (1.2 +/- 0.6, P < 0.0001), with no large follicles from Day 9. The number of medium follicles (4-5 mm in size) also decrease during the period of treatment (4.2 +/- 0.7 to 1.0 +/- 0.6, P < 0.0001), leading to a significant decrease in inhibin A levels when compared to the control (143.7 +/- 31.3 pg/ml versus 65.2 +/- 19.1 pg/ml, P < 0.00005). In contrast, the number of small follicles (2-3 mm) increased in treated goats from Day 4 of treatment (9.6 +/- 2.9 to 20.2 +/- 6.3, P < 0.005). Such data indicate that GnRH antagonist reduced plasma levels of FSH and LH with suppression of the growth of large dominant ovarian follicles and a two-fold increase in number of smaller follicles. The results confirm that GnRH antagonist treatment can be used in goats to control gonadotrophin secretion and ovarian follicle growth in superovulatory regimes.

The Booroola (FecB) phenotype is associated with a mutation in the bone morphogenetic protein (BMP) receptor 1B. The BMP action is important during development; surprisingly the only differences so far observed in adult animals are restricted to the ovaries where precocious development of the antral follicles and increased ovulation rate of mutant ewes is observed. The internal organs of 17 ewes homozygous for the mutation (BB) and 18 wild-type ewes (++) were macroscopically examined and weighed. No macroscopic differences were found, and the weight of the heart, liver, lungs, kidneys, and spleen were similar for both genotypes (P > 0.05). In contrast, the adrenals of BB ewes were lighter than those of ++ ewes (P < 0.05). The effect of the mutation on the adrenal function of cortisol secretion was measured at basal level and after an adrenocorticotrophic hormone challenge, before and after dexamethasone suppression. The Booroola mutation had no effect (P > 0.05) in any of these conditions. These findings indicate that the Booroola mutation also affects the size of the adrenal glands and suggest that the mutated gene could be important in the development of other organs in addition to the ovary. However, in the mutant ewes the function of the adrenal glands is not compromised or it is compensated.

To test whether inhibin A assays can be used for the prediction of yields in embryo programmes in goats, 50 does were treated with 45 mg FGA sponges (Chronogest) for 16 days plus a single dose of 100 microg i.m. cloprostenol on Day 14, just before the start of administration of eight doses of 1.25 ml of Ovagen twice daily for 4 days. At first FSH injection, the number and size of all follicles > or =2 mm was assessed by transrectal ultrasound and plasma inhibin A levels were measured by specific dimeric assay. There was a positive correlation between number of follicles > or =6 mm (8.8 +/- 0.5) and inhibin A levels at first FSH dose (193.2 +/- 14.5 pg/ml, P<0.05). The mean number of corpora lutea on Day 7 after sponge removal was related to the total number of follicles with a diameter of 2-6 mm at the onset of the FSH treatment (15.3 +/- 0.7, P<0.05). The total number of embryos recovered was related to the number of follicles with 4-6 mm in size (6.2 +/- 0.5, P<0.05) and to the inhibin A levels at first FSH dose (P<0.05). These results suggest that follicles > or =4 mm are the source of inhibin prior to FSH stimulation and are the main source of oocytes resulting in the number of viable embryos recovered after a superovulatory treatment. Hence, the response to superovulatory treatments in goats in terms of the number of embryos can be predicted from either the population of follicles determined by ultrasound or the plasma inhibin A levels at start of the superovulatory FSH treatment.

The prolificacy variation in sheep makes it an excellent animal model to understand the mechanisms regulating ovulation rate. Identification of mutations responsible for the increased prolificacy of the Inverdale, Booroola, Javanese, Cambridge and Belclare sheep open new avenues of investigation for the paracrine control of folliculogenesis. To date, all known mutations are in genes from ligands or receptors of the transforming growth factor beta superfamily, and point to the bone morphogenetic protein family of peptides as local regulators of ovarian follicle growth. The mechanism of action of the mutated genes is not fully understood, but results in the ovulation of a higher number of follicles with smaller diameter and fewer granulosa cells than that of the wildtype, thus speeding the differentiation of ovulatory follicles. Comparisons of the performance of Booroola-crossed flocks in different countries showed that carriers of the prolificacy mutation have higher ewe productivity but also higher perinatal mortality and lighter weight lambs. Their economic impact on the sheep industry depends on farm environment and management. Nevertheless, the diagnostic tests now available to identify the genetic mutations resulting in increased ovulation rate, will simplify the introduction of these mutations and their monitoring in flocks for research and commercial purposes.

This review offers an overview of the basic characteristics of in vivo embryo technologies, their current status, the main findings and the advances gained in recent years, and the outstanding subjects for increasing their efficiency. The use of superovulation and embryo transfer procedures remains affected by a high variability in the ovulatory response to hormonal treatment and by a low and variable number of transferable embryos and offspring obtained. This variability has been classically identified with both extrinsic (source, purity of gonadotrophins and protocol of administration) and intrinsic factors (breed, age, nutrition and reproductive status), which are reviewed in this paper. However, emerging data indicate that the main causes of variability are related to endocrine and ovarian factors, and so the number of studies and procedures addressing a better understanding and control of these factors may be increased in the future. The accomplishment of this objective, the improvement of procedures for embryo conservation and for the selection and management of recipient females, will allow further development and application of this technology.

The Booroola phenotype is associated with a point mutation in the kinase domain of the bone morphogenetic protein receptor 1 B (BMPR1 B), and is characterized by 'precocious' differentiation of ovarian follicles, leading to the production of large numbers of ovulatory follicles that are smaller in diameter than wild-type follicles. These smaller follicles attain differentiation markers, such as expression of mRNA for P450 aromatase and inhibin-betaA subunit, granulosa cell LH receptors and aromatase activity, earlier than follicles from wild-type ewes. However, the preovulatory follicles from mutant ewes collectively secrete similar quantities of oestradiol, androstenedione and inhibin A in exactly the same pattern as wild-type ewes, which result in similar concentrations of FSH. The available evidence strongly indicates that the Booroola mutation exerts its action at the ovary rather than by altering gonadotrophin secretion. The bone morphogenetic protein (BMP) receptors and putative ligands are ubiquitously expressed within the ovary and BMPs seem to be involved in the paracrine regulation of FSH action. Thus, if the mutation is causing a reduction in BMPR1 B signalling, it may act on an inhibitor of follicle differentiation. Further research in this area will concentrate on the elucidation of the natural ligands for BMPR1 B at different stages of follicle development and examine the effect of BMPR1 B mutation on the downstream signalling cascade.

Objectives of this study were to characterize patterns of follicular development in sheep superovulated with purified follicle stimulating hormone (FSH) (OVAGEN, ICP, Auckland, New Zealand) and to determine its influence on preovulatory events (onset of the oestrus behaviour and timing of the preovulatory luteinizing hormone surge) and ovarian response (ovulation rate and embryo yield). Number and size of all >/= 23 mm follicles from the first FSH injection to withdrawal of progestagen sponges was determined by transrectal ultrasonography just prior to every FSH injection in nine Manchega ewes superovulated with eight decreasing doses (ml) (1.5 x 3, 1.25 x 2 and 1 x 3) of OVAGEN injected twice daily from 60 h before to 24 h after the withdrawal of 40 mg fluorogestone acetate sponges. Oestrous detection and jugular blood sampling for LH radioimmunoassay were performed every 3 h from 14 to 53 h after sponge removal and ovulation rate and number of embryos were determined 4 days after progestagen withdrawal. Administration of OVAGEN induced a significant rise (p < 0.0005) in the number of follicles >/= 4 mm in size because of an increased growth in size of follicles from the first FSH injection to sponge removal, an increase in the number of newly detected follicles from 12 to 36 h of the first FSH dose (p < 0.005) and a decrease in regression rate from 24 h (p < 0.001). The number of follicles 2-3 mm in size at first FSH dose (10.4 +/- 1.5) was positively correlated with the number of >/= 4 mm follicles at 0 h (19.0 +/- 2.7, p < 0.01). A higher number of >/= 4 mm follicles at 0 h was related with an earlier appearance of oestrus (31.5 +/- 1.5 h, p = 0.08) and LH surge (45.0 +/- 2.3 h, p < 0.005), and a higher ovulation rate (18.2 +/- 3.8, p < 0.005). On the other hand, the rate of embryo recovery was decreased in ewes with earlier preovulatory LH peaks (p < 0.005), with a shorter interval between oestrus and LH peak (p < 0.05).

Variability in superovulatory response to FSH stimulation is common to most mammals and imposes practical problems for assisted reproduction. In sheep, we have studied if this response is related to the ovarian follicular population and activity before the stimulation. During the breeding season, 30 ewes were treated with 40 mg FGA sponges for 14 days and 125 microg cloprostenol injection on Day 12, considering Day 0 as the day of progestagen insertion. Superovulatory response was induced with two different FSH regimes using the same total dose (8.8 mg), administered twice daily from 60 h before to 24 h after progestagen withdrawal. At the first FSH injection, all follicles > or = 2 mm were observed by transrectal ultrasonography and plasma FSH and inhibin A levels were determined. The number of corpora lutea and the number of and viability of recovered embryos in response to the treatment were determined on Day 7 after sponge withdrawal. No significant differences were found between treatments. The total mean number of corpora lutea (11.5 +/- 1.2) and recovered embryos (7.9 +/- 1.1) were positively correlated (P < 0.05 and <0.01, respectively) with the number of small antral follicles (2-3 mm: 9.2 +/- 0.7) and inhibin A concentration (240 +/- 18 pg/ml; P < 0.05 for corpora lutea and P < 0.005 for recovered embryos) observed at the onset of the superovulatory treatment, which was also positively correlated with the number of viable embryos (5.8 +/- 0.9, P < 0.005). In 18 ewes with follicles > or = 6 mm prior to FSH treatment, the ovulation rate was unaffected but the number of embryos (6.1 +/- 0.9 versus 11.6 +/- 2; P < 0.05) and their viability (4.5 +/- 0.8 versus 8.5 +/- 2; P < 0.05) was reduced. The lower number of embryos produced when a large follicle is present suggest that a proportion of the smaller follicles are in early stages of atresia and the developmental competence of their oocyte is compromised.