ABSTRACT
This qualitative narrative study aimed to examine how a family of origin influenced the professional self of Colombian students pursuing a profession in family therapy. Few research studies have investigated the development of self among native Spanish speakers from Colombia. Undergraduate and graduate students enrolled in clinical psychology and family therapy internships (N = 16) from the Colombian cities of Medellín, Pereira, and Santa Marta participated in this study. Data collection using personal diaries, family albums, autobiographies, and in-depth interviews served as sources for narrative analysis. Two themes and four subthemes representing positive and negative aspects of influences of families of origin helped organize the findings. Student experiences illustrate abilities, skills, and potentialities in the narratives. Implications for clinical training and future research recommendations highlight a path forward.
Subject(s)
Family Therapy , Students , Humans , Colombia , Qualitative Research , Clinical CompetenceABSTRACT
Equine intracytoplasmic sperm injection (ICSI) is being used clinically for foal production, but little information is available on factors affecting the efficiency of this procedure. We examined factors that may influence blastocyst development when ICSI is performed clinically, i.e., on oocytes recovered from live mares by transvaginal ultrasound-guided follicle aspiration (TVA), using sperm from the stallion of the client's choice. In a clinical setting, there may be a delay from the time of TVA to isolation of oocytes from the aspirated fluid. In a preliminary study, oocytes from fluid held for 1.5 h at ambient temperature (26°C-33°C) yielded 32% blastocysts; however, in experiment 1, fluid held at 32 °C for 2 h after aspiration yielded 16% blastocysts versus 23% for aspirates processed immediately. Performing TVA/ICSI throughout the year would increase production from valuable mares, but efficiency during the nonbreeding season is unknown. In addition, to reduce the possibility of infection after TVA, administration of antibiotics to the mare before TVA is indicated; however, these could affect oocyte quality. In experiment 2, follicle numbers at the time of TVA were significantly higher in December to January than for the same mares during the breeding season. Oocyte recovery rates on TVA were 60% to 66% and the blastocyst rate was 18%. An equivalent blastocyst rate (18%) was achieved after administration of ampicillin and gentamicin to mares before TVA. In experiment 3, we verified that stallion differences exist in rates of cleavage after ICSI with motile sperm. In sperm from a low-performing stallion, density-gradient centrifugation followed by swim-up was associated with significantly higher rates of cleavage (45% vs. 18%) and blastocyst development (14% vs. 0%) than those for density gradient alone. In experiment 4, parthenogenetic activation with ionomycin and 6-dimethylaminopurine yielded 40% blastocysts. Frozen-thawed sperm that were immotile after nitrogen tank failure did not produce blastocysts; exogenous activation after ICSI increased cleavage rate but did not yield blastocysts. These studies provide information on factors that may affect in vitro blastocyst formation after equine ICSI as it is performed in a clinical program.
Subject(s)
Blastocyst/physiology , Embryo Culture Techniques/veterinary , Horses/embryology , Sperm Injections, Intracytoplasmic/veterinary , Animals , Female , In Vitro Oocyte Maturation Techniques/veterinary , Male , Spermatozoa/physiologyABSTRACT
Equine embryos develop in vitro in the presence of high glucose concentrations, but little is known about their requirements for development. We evaluated the effect of glucose concentrations in medium on blastocyst development after ICSI. In experiment 1, there were no significant differences in rates of blastocyst formation among embryos cultured in our standard medium (DMEM/F-12), which contained >16âmM glucose, and those cultured in a minimal-glucose embryo culture medium (<1âmM; Global medium, GB), with either 0 added glucose for the first 5 days, then 20âmM (0-20) or 20âmM for the entire culture period (20-20). In experiment 2, there were no significant differences in the rates of blastocyst development (31-46%) for embryos cultured in four glucose treatments in GB (0-10, 0-20, 5-10, or 5-20). Blastocysts were evaluated by immunofluorescence for lineage-specific markers. All cells stained positively for POU5F1. An inner cluster of cells was identified that included presumptive primitive endoderm cells (GATA6-positive) and presumptive epiblast (EPI) cells. The 5-20 treatment resulted in a significantly lower number of presumptive EPI-lineage cells than the 0-20 treatment did. GATA6-positive cells appeared to be allocated to the primitive endoderm independent of the formation of an inner cell mass, as was previously hypothesized for equine embryos. These data demonstrate that equine blastocyst development is not dependent on high glucose concentrations during early culture; rather, environmental glucose may affect cell allocation. They also present the first analysis of cell lineage allocation in in vitro-fertilized equine blastocysts. These findings expand our understanding of the factors that affect embryo development in the horse.
Subject(s)
Blastocyst/drug effects , Cell Lineage/drug effects , Cell Lineage/physiology , Embryonic Development/drug effects , Glucose/administration & dosage , Animals , Blastocyst/cytology , Embryonic Development/physiology , HorsesABSTRACT
In nuclear transfer (NT), exposure of donor cell chromatin to the ooplast cytoplasm may aid reprogramming; however, the length of exposure feasible is limited by the developmental life span of the oocyte. We examined the effect of duration of nucleus-cytoplasmic exposure before activation and of in vitro maturation (IVM) in equine NT. In experiment 1, 24 h IVM and a delay of 2, 5, or 8 h between reconstruction and activation yielded 4%, 15%, and 11% blastocysts, respectively. In experiment 2, a 5-h activation delay yielded 17% and 22% blastocysts with two donor cell lines. In experiment 3, using a 5-h activation delay, the blastocyst rate was significantly higher using oocytes after 20 h IVM than after 24 h IVM; however, only 28% of oocytes were in metaphase II (MII) at 20 h. In experiment 4, oocytes were denuded of cumulus at 20 h, and those in metaphase I (MI) were returned to culture for 3 h (20+3H treatment); blastocyst rates were 30% and 27%, respectively (8-h and 5-h delay to activation, respectively). Four live foals resulted from the transfer of 17 blastocysts (24%) produced using MII oocytes and a 5- or 8-h activation delay. Use of equine oocytes immediately after reaching MII, combined with a longer delay from reconstruction to activation, increased developmental competence after equine NT.
Subject(s)
Blastocyst/cytology , Embryonic Development , Nuclear Transfer Techniques/veterinary , Oocytes/cytology , Animals , Embryo Transfer/veterinary , Female , Fertilization in Vitro/veterinary , Horses , Metaphase , Time FactorsABSTRACT
In vitro fertilization does not occur readily in the horse. This may be related to failure of equine sperm to initiate hyperactivated motility, as treating with procaine to induce hyperactivation increases fertilization rates. In mice, hyperactivated motility requires a sperm-specific pH-gated calcium channel (CatSper); therefore, we investigated this channel in equine sperm. Motility was assessed by computer-assisted sperm motility analysis and changes in intracellular pH and calcium were assessed using fluorescent probes. Increasing intracellular pH induced a rise in intracellular calcium, which was inhibited by the known CatSper blocker mibefradil, supporting the presence of a pH-gated calcium channel, presumably CatSper. Hyperactivation was associated with moderately increased intracellular pH, but appeared inversely related to increases in intracellular calcium. In calcium-deficient medium, high-pH treatment induced motility loss, consistent with influx of sodium through open CatSper channels in the absence of environmental calcium. However, sperm treated with procaine in calcium-deficient medium both maintained motility and underwent hyperactivation, suggesting that procaine did not act via opening of the CatSper channel. CATSPER1 mRNA was identified in equine sperm by PCR, and CATSPER1 protein was localized to the principal piece on immunocytochemistry. Analysis of the predicted equine CATSPER1 protein revealed species-specific differences in structure in the pH-sensor region. We conclude that the CatSper channel is present in equine sperm but that the relationship of hyperactivated motility to calcium influx is weak. Procaine does not appear to act via CatSper in equine sperm, and its initial hyperactivating action is not dependent upon external calcium influx.
Subject(s)
Calcium Channels/physiology , Calcium/metabolism , Horses/physiology , Sperm Motility/genetics , Spermatozoa/physiology , Animals , Calcium Channels/genetics , Calcium Signaling/genetics , Horses/genetics , Hydrogen-Ion Concentration , Intracellular Space/metabolism , Kinetics , Male , RNA, Messenger/metabolism , Spermatozoa/metabolism , Tissue DistributionABSTRACT
The presence of heterogenous mitochondria from the host ooplast affects the acceptance of offspring obtained by somatic cell nuclear transfer. This might be avoided by obtaining oocytes from selected females, but is then complicated by low numbers of available oocytes. We examined the efficiency of equine somatic cell nuclear transfer using oocytes recovered by transvaginal aspiration of immature follicles from 11 mares. Use of metaphase I oocytes as cytoplasts and of scriptaid (a histone deacetylase inhibitor) treatment during oocyte activation were evaluated to determine if these approaches would increase blastocyst production. In experiment 1, blastocyst development was 0/14 for metaphase I oocytes and 4/103 (4%) for metaphase II oocytes. Three blastocysts were transferred to recipient mares, resulting in two pregnancies and one live foal, which died shortly after birth. In experiment 2, blastocyst development was 2/47 (4%) for control oocytes and 1/83 (1%) for scriptaid-treated oocytes. No foals were born from two blastocysts transferred in the control group. The blastocyst from the scriptaid treatment resulted in birth of a live foal. In conclusion, this is apparently the first report of production of a viable cloned foal from oocytes collected from immature follicles of live mares, supporting the possibility of cloning using oocytes from selected mares.
Subject(s)
Cloning, Organism/veterinary , Horses/physiology , Nuclear Transfer Techniques/veterinary , Animals , Cloning, Organism/methods , Embryo Transfer/veterinary , Embryonic Development , Female , Pregnancy , Pregnancy RateABSTRACT
OBJECTIVE: To evaluate the efficiency of foal production following intracytoplasmic sperm injection (ICSI) and blastocyst culture of oocytes from mares that died or were euthanized under field conditions. DESIGN: Prospective case series. ANIMALS: 16 mares (age, 3 to 19 years) that died or were euthanized for various causes. PROCEDURES: Ovaries were collected immediately before euthanasia (n = 10) or after death (6). Ovaries were transported to the laboratory for oocyte recovery (15 mares), or oocytes were recovered at a remote location and shipped to the laboratory (1). Oocytes underwent ICSI, and presumptive zygotes were cultured for 7 to 10 days. Blastocysts were shipped to embryo transfer facilities for transcervical transfer to recipient mares. RESULTS: Ovaries were processed 30 minutes to 12 hours (mean ± SD, 4.6 ± 3.3 hours) after mares' deaths. A mean of 14.1 ± 8.6 oocytes/mare were cultured, and 110 of 225 (49%) matured. Twenty-one blastocysts developed after ICSI and were transferred to recipient mares. Thirteen pregnancies were established; 10 healthy foals were produced from 6 donor mares. The number of blastocysts produced per mare and number of live foals produced per mare were significantly correlated with the number of oocytes recovered. CONCLUSIONS AND CLINICAL RELEVANCE: Foals were produced from mares after death or euthanasia under field conditions. Proportions of foals born overall (10 foals/16 mares) and mares from which ≥ 1 foal was produced (6/16) were greater than those reported following recovery and oviductal transfer of oocytes to inseminated recipients after death of donor mares under field conditions.
Subject(s)
Blastocyst/physiology , Embryo Culture Techniques/veterinary , Horses/physiology , Oocytes/physiology , Ovary/physiology , Sperm Injections, Intracytoplasmic/veterinary , Animals , Euthanasia, Animal , FemaleABSTRACT
Objective: study the influence of time of exposure to hypobaric hypoxia and pulmonary weight:body weight ratio (PW:CW) on the development of pulmonary hypertension (PH) in Cobb Vantress 500® broiler chickens. Methods: five groups of birds were transfered from a relative normoxic environment (336 masl) to a hypobaric hypoxic one (2638 masl) at the following ages (in days): 6, 10, 15, 20 and 25. An additional group remained at 2638 masl from day 1. All birds were slaughtered at day 49 and the incidence of PH was assessed, as well as PW:CW. Results: no statistical association between age of exposure or PW:CW and PH was found (p<0.05). PH incidence was higher than those previously reported in similar studies. Since there was a strict temperature control in the present study, this factor can be excluded as a possible cause of PH. Further experiments should be conducted to assess the relative importance of other contributing factors in the occurrence of PAH.
Objetivo: el objetivo de este estudio fue evaluar la posible injerencia del desarrollo pulmonar y el tiempo de permanencia sobre la incidencia de hipertensión arterial pulmonar (HAP) en pollos de engorde, de la estirpe Cobb Vantress 500®. Métodos: Se trasladaron cinco grupos de aves desde un sitio con condiciones de normoxia relativa (336 metros sobre el nivel del mar msnm) a uno con condiciones de hipoxia hipobárica (2638 msnm) en las siguientes edades: 6, 10, 15, 20 y 25 días. Hubo un grupo de animales que permaneció desde el día 1 bajo hipoxia hipobárica. Todos los animales fueron sacrificados a los 49 días de edad. Entonces, se calculó la incidencia de HAP y la relación peso pulmonar : peso corporal (PP:PC) en pollos sanos y en aquellos que tuvieron HAP. Resultados: No hubo asociación estadística entre la edad de exposición a condiciones de hipoxia o de la relación PP:PC y la presentación de la enfermedad (p<0.05). La incidencia de HAP fue más alta que en estudios anteriores llevados a cabo con la misma estirpe y en condiciones comparables a las de la presente investigación. Dado que hubo un control de la temperatura ambiental en el sitio de alojamiento de los pollos, ese factor reconocido como desencadenante de HAP, puede ser excluido en las condiciones del presente estudio. Es deseable que se diseñen experimentos para valorar la importancia relativa de otros factores coadyuvantes en la presentación de HAP.
Objetivo: Com o propósito de avaliar a possível ingerência do desenvolvimento do pulmão e do tempo sobre a incidência de hipertensão arterial pulmonar (HAP) em frangos de corte da linha genética Cobb Vantress 500®. Métodos: As aves foram mudadas em 5 grupos de um local com condições de normoxia relativa (336 m acima do nível do mar) para uma com condições de hipoxia hipobárica (2638 m) nas idades seguintes: 6, 10, 15, 20 e 25 dias. Houve um grupo de animais que permaneceram desde o primeiro dia em condições de hipoxia. Todos os animais foram sacrificados aos 49 dias de idade. Posteriormente foi calculada a incidência de HAP e peso pulmonar: peso corporal (PP: PC) nos frangos e naqueles que tinham HAP. Resultados: Não houve associação estatística entre a idade de exposição à hipoxia ou a relação PP: PC e a apresentação da doença (p<0.05). A incidência de HAP foi maior do que nos estudos anteriores realizados com a mesma linha genética e em condições comparáveis às do presente estudo. Como houve um controle de temperatura do local nos primeiros dias de idade, que é reconhecido como um fator importante para a HAP, pode ser excluído nas condições deste estudo. É desejável elaborar experimentos para avaliar a importância relativa de outros fatores que contribuem para a apresentação da HAP.
ABSTRACT
Se presenta un caso de infección zigomicótica en un paciente de 52 años con diagnóstico previo de linfoma no Hodgkin. Aunque la infección micótica por Absidia sp. es favorecida en este caso por condiciones subyacentes como leucopenia severa con deficiencia de granulocitos y tratamiento quimioterapéutico agresivo, la forma de presentación rinoorbitaria sin compromiso cerebral al igual que la ausencia de otras características clínicas clásicas de este tipo de compromiso es notable. Se observan en este caso manifestaciones pulmonares concomitantes pero desafortunadamente, no se llega a establecer su etiología aunque se sospecha fuertemente una diseminación micótica.