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PubMed1997

Interaction between triiodothyronine and ovarian steroid hormones on the regulation of the release of thyrotropin and thyrotropin-releasing hormone in vitro.

In vivo and in vitro experiments were designed to examine [1] the effect of triiodothyronine (T3) and/or ovarian steroids on the spontaneous and thyrotropin-releasing hormone (TRH)-stimulated release of thyrotropin (TSH) by the anterior pituitary gland (AP) in vitro; and [2] the in vivo effects of T3 and ovarian steroids on TRH-release in vitro. In the experiment 1, ovariectomized-thyroidectomized (Ovx-Tx) rats were injected with triiodothyronine (T3, 2 micrograms/kg), estradiol benzoate (EB, 25 micrograms/kg), progesterone (P, 10 mg/kg), T3 plus EB, T3 plus P, EB plus P, or T3 plus EB and P for 6 days before decapitation. The AP was incubated with Locke's medium, challenged with TRH (30 nM), recovered and then with T3 (10 nM) only or with T3+TRH, 30 min for each interval. Mediobasal hypothalami (MBHs) were challenged with high potassium (60 mM) for 30 min. In the experiment 2, the APs of Ovx-Tx rats were enzymatically dispersed and the AP cells were pretreated with or without EB (0-6 nM) for 72 h, and further with T3 (10 nM) for 24 h, followed by an incubation for 30 min with TRH (0-100 nM). The spontaneous and TRH-induced release of TSH in vitro from rat APs, and pituitary TSH content were increased by T3, or T3 plus P as compared with the animals injected with vehicle, or P alone. EB inhibits the effect of T3 on TSH release in vitro. Application of T3 in vitro prevented the release of TSH in response to TRH. EB dose-dependently relieved the inhibitory effect of T3 on TRH-induced TSH release in vitro. TRH release from MBH was increased by EB and inhibited by T3 or P. EB prevented the inhibitory effect of T3 on TRH release. P plus T3 potentiated the stimulatory effects of EB on TRH release. These results suggest that [1] the reduction of the concentration of plasma TSH by T3 is at least in part due to the inhibitory effects of T3 on TRH release from mediobasal hypothalamus, and TSH release in response to TRH, [2] the increased content and release of TSH from rat AP tissue by T3 via an in vivo effect may be involved in a short feedback loop of TSH on TRH release, and [3] ovarian steroid hormones play an inhibitory role in regulating T3 effects on the release of TSH and TRH.

Animals↗

PubMed1998

Effects of in vitro versus in vivo red cell labeling on image quality in gastrointestinal bleeding studies.

OBJECTIVE: Both in vivo and in vitro red cell labeling methods are available for performing gastrointestinal bleeding studies. While in vitro labeling has been shown to result in higher binding efficiency, no comparison of clinical image quality has been reported between those techniques. This study compares in vivo and in vitro methods using both subjective and objective measurements of image quality. METHODS: A consecutive series of gastrointestinal bleeding studies performed on 23 patients using in vivo labeling was compared to a series of 23 studies using in vitro labeling. Images at 30 min postinjection were randomized and analyzed by two observers. Subjective evaluation of image quality, as well as renal activity, was based on a comparison of femoral vein and inferior vena cava activity to adjacent background using numerical scores with a scale of 0-3. Image quality using the subjective scores was further classified as acceptable and poor. Target-to-background measurements of femoral vein to adjacent soft tissue in the thigh and inferior vena cava to adjacent abdominal background also were made. RESULTS: In vitro label subjective image quality was acceptable in 87% and 91% of cases for vascular and renal activity, respectively, but only 35% and 52% for in vivo labeling. In vitro label target-to-background ratios were significantly better than in vivo label for the femoral vein and inferior vena cava, as well as for the subjective assessment of vascular image quality, but not for renal activity. CONCLUSION: In vitro red cell labeling improves clinical image quality as compared with in vivo labeling. Both subjective and objective measurements of image quality are useful for comparing the results of labeling methods.

Abdomen↗

PubMed1998

Prednisolone resistance in childhood acute lymphoblastic leukemia: vitro-vivo correlations and cross-resistance to other drugs.

As an important determinant of response to chemotherapy, accurate measurement of cellular drug resistance may provide clinically relevant information. Our objectives in this study were to determine the relationship between in vitro resistance to prednisolone (PRD) measured with the colorimetric methyl-thiazol-tetrazolium (MTT) assay, and (1) short-term clinical response to systemic PRD monotherapy, (2) long-term clinical outcome after combination chemotherapy within all patients and within the subgroups of clinical good and poor responders to PRD, and (3) in vitro resistance to 12 other drugs in 166 children with newly diagnosed acute lymphoblastic leukemia (ALL). The 12 clinical poor PRD responders had ALL cells that were median 88-fold more in vitro resistant to PRD than 131 good responders (P = .013). Within all patients, increased in vitro resistance to PRD predicted a significantly worse long-term clinical outcome, at analyses with and without stratification for clinical PRD response, and at multivariate analysis (P </= .001). Within both the clinical good and poor responder subgroups, increased in vitro resistance to PRD was associated with a worse outcome, which was significant within the group of clinical good responders (P < .001). LC50 values, ie, lethal concentrations to 50% of ALL cells, for PRD and each other drug correlated significantly with those of all other 12 drugs, with an average correlation coefficient of 0.44 (standard deviation 0.05). The highest correlations were found between structurally related drugs. In conclusion, in vitro resistance to PRD was significantly related to the short-term and long-term clinical response to chemotherapy, the latter also within the subgroup of clinical good responders to PRD. There was a more general in vitro cross-resistance between anticancer drugs in childhood ALL, although drug-specific activities were recognized.

Adolescent↗

PubMed1989

Clinical significance of crossed in vitro sperm-cervical mucus penetration test in infertility investigation.

To evaluate the clinical significance of in vivo and in vitro testing of sperm ability to penetrate cervical mucus (CM), postcoital testing (PCT) and in vitro sperm-cervical mucus penetration testing were compared in a prospective study. Both in vivo and in vitro tests were standardized and performed after an oral course of estrogen therapy. Crossed in vitro sperm-cervical mucus penetration test, evaluated in 277 couples with CM of patients' wives and additionally with CM and semen of fertile donors, revealed that the male factor contributed to a significantly higher extent to deficient sperm-mucus interaction than the cervical factor. The overall pregnancy rate after 6 months was 23% (64/277). Whereas the outcome of PCT did not significantly predict subsequent fertility (PCT good pregnancy rate 24%/PCT poor 20%), significant differences were found for the sperm-cervical mucus penetration test with CM of patients' wives (pregnancy rate, 30.5% versus 8.5%) and for in vitro testing with donors' CM, but not for the mucus penetration test with donors' spermatozoa. Routine sperm analysis did not prove to be of prognostic value for a subsequent pregnancy. The results suggest that the in vitro sperm-cervical mucus penetration test is a good parameter of sperm function and, in particular, when performed as a cross-matching penetrability test, a valuable adjunct to PCT with regard to fertility prognosis.

Cervix Uteri↗

PubMed1995

Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching.

Immature oocyte recovery followed by in-vitro oocyte maturation and in-vitro fertilization is a promising new technology for the treatment of human infertility. The technology is attractive to potential oocyte donors and infertile couples because of its reduced treatment intervention. Immature oocytes were recovered by ultrasound-guided transvaginal follicular aspiration. Oocytes were matured in vitro for 36-48 h followed by intracytoplasmic sperm injection (ICSI). Embryos were cultured in vitro for 3 or 5 days before replacement. Assisted hatching was performed on a day 5 blastocyst stage embryo. Embryo and uterine synchrony were potentially enhanced by luteinization of the dominant follicle at the time of immature oocyte recovery. Mature oocyte and embryo production from immature oocyte recovery were similar to the previous IVF results of the patients. A blastocyst stage embryo, produced as a result of in-vitro maturation, ICSI, in-vitro culture and assisted hatching, resulted in the birth of a healthy baby girl at 39 weeks of gestation.

Blastocyst↗

PubMed1995

Polyamines may increase the percentage of in-vitro fertilized murine oocytes that develop into blastocysts.

This study was designed to investigate whether the polyamines spermine, spermidine and putrescine can prevent the arrest of development of in-vitro fertilized murine oocytes from CD1 strain mice. Development in M16 or Brinster's medium with or without polyamine supplementation was assessed and the ability of blastocysts to attach and grow out on laminin was compared with in-vivo-generated embryo blastocysts. Oocytes from old (20 weeks) females showed lower rates of fertilization and subsequent development in comparison with oocytes from young (6-8 weeks old) females, but in both cases < 5% of in-vitro fertilized oocytes achieved the blastocyst stage. In-vitro fertilized young oocytes showed enhanced development to the blastocyst stage with the addition of the polyamine putrescine, although lesser enhancement was seen with spermine and spermidine; this effect was only evident when the oocytes were grown in Brinster's medium, reflecting the need for a particular set of culture conditions for growth-enhancing agents to be effective. Blastocysts generated in putrescine-supplemented Brinster's medium were able to implant on laminin-coated surfaces in vitro and to respond to the cytokines colony-stimulating factor-1, epidermal growth factor and platelet-derived growth factor with increased trophoblast outgrowth to a greater degree than in-vivo-generated blastocysts. Murine in-vitro fertilized embryo development was not enhanced by co-culture with Vero cells. Putrescine was the most effective polyamine, enhancing the in-vitro fertilized oocyte development to blastocysts in a permissive environment.

Animals↗

PubMed1999

Monoamine oxidase A inhibition by fluoxetine: an in vitro and in vivo study.

Monoamine oxidase A (MAO-A) inhibition was investigated both in vitro and in vivo in rat brains by using the radioligand, 18F-fluoroclorgyline (N-[3-(2',4'-dichlorophenoxy)-2-18F-fluoropropyl]-N-methylpropa rgylamine). In vitro binding affinities of six compounds, clorgyline, Ro 41-1049, deprenyl, fluoxetine, norfluoxetine and citalopram, were studied. Fluoxetine and norfluoxetine showed in vitro affinities of 36.5 and 68 microM for MAO-A, respectively. Fluoxetine and norfluoxetine also significantly inhibited (more than 20%) the binding of the radioligand in vivo while citalopram and deprenyl showed very poor affinities in vitro for MAO-A and had no effect in vivo. The in vivo effects of the various drugs were directly comparable to their in vitro affinities for binding to MAO-A as seen in the correlation plot of percent control in vivo binding of 18F-fluoroclorgyline and binding affinity, -log IC50 (R2 = 0.979). An acute dose of 20 mg/kg of fluoxetine inhibited binding of 18F-fluoroclorgyline by more than 20%, while lower doses had some significant effects. These results provide evidence on the in vitro and in vivo inhibition of monoamine oxidase A by fluoxetine.

Animals↗

PubMed1998

The promotory effect of growth hormone on the developmental competence of in vitro matured bovine oocytes is due to improved cytoplasmic maturation.

In a previous study we have shown that the addition of growth hormone (GH) during in vitro maturation accelerates nuclear maturation, induces cumulus expansion, and promotes subsequent cleavage and embryonic development. The aim of this study was to investigate whether the promotory effect of GH on subsequent cleavage and blastocyst formation is due to an improved fertilization and whether this effect is caused by an improved cytoplasmic maturation of the oocyte. Therefore, bovine cumulus oocyte complexes (COCs) were cultured for 22 hours in M199 supplemented with 100 ng/ml bovine GH (NIH-GH-B18). Subsequently the COCs were fertilized in vitro. Cultures without GH served as controls. To verify whether the promoted fertilization is caused by the effect of GH on cumulus expansion or oocyte maturation, cumulus cells were removed from the oocytes after in vitro maturation (IVM) and denuded MII oocytes were selected and fertilized in vitro. Both IVM and in vitro fertilization (IVF) were performed at 39 degrees C in a humidified atmosphere with 5% CO2 in air. At 18 hours after the onset of fertilization, the nuclear stage of the oocytes was assessed using 4,6-diamino-2-phenylindole (DAPI) staining. Oocytes with either an metaphase I (MI) or MII nuclear stage and without penetrated sperm head were considered unfertilized; oocytes with two pronuclei, zygotes, and cleaved embryos were considered normally fertilized; and oocytes with more than two pronuclei were considered polyspermic. To evaluate cytoplasmic maturation, the distribution of cortical granules 22 hours after the onset of IVM, and sperm aster formation 8 hours after the onset of fertilization were assessed. In addition, to assess the sperm-binding capacity, COCs were fertilized in vitro, and 1 hour after the onset of fertilization the number of spermatozoa bound to the oocytes was counted. The addition of GH during IVM significantly (P < 0.001) enhanced the proportion of normal fertilized oocytes. Removal of the cumulus cells prior to fertilization and selection of the MII oocytes did not eliminate the positive effect of GH on fertilization. No effect of GH on the sperm-binding capacity of the oocyte was observed. In addition, GH supplementation during IVM significantly (P < 0.001) enhanced the migration of cortical granules and sperm aster formation. It can be concluded that the promotory effect of GH on the developmental competence of the oocyte is due to a higher fertilization rate as a consequence of an improved cytoplasmic maturation.

Animals↗

PubMedDate not supplied

Effect of laundering on ability of glove fabrics to decrease the penetration of organophosphate insecticides through in vitro epidermal systems.

Two knit glove fabrics, one of 100% cotton and one of 100% polypropylene, were examined for their capability to decrease the penetration of the organophosphate insecticides (OPs), azinphos-methyl and paraoxon after 4 h at field concentrations (3000 and 15 ppm, respectively) through an in vitro epidermal system (Skin2, Advanced Tissue Systems, LaJolla, CA). The glove fabrics were examined under three different conditions of use: new, after they had been abraded and after they had been abraded and then laundered. New and laundered cotton fabric was also examined for its capability to decrease the penetration of azinphos-methyl through another in vitro epidermal system (Epiderm, MatTek Corp., Ashland, MA), after 4 and 24 h of exposure. Capability of the media under the in vitro epidermal systems to inhibit brain acetylcholinesterase (AChE) was used as the indicator of penetration. Results were compared to OP-caused inhibitions seen in media under the fabric alone and in media under the in vitro epidermal systems alone. Incubations of azinphos-methyl suspensions and the in vitro epidermal systems covered with fabric indicated that both the epidermal cells and fabric provided protection against AChE inhibition caused by this OP and that the protective effects were additive, whether measured after 4 or 24 h of exposure. Therefore, neither laundering nor abrasion followed by laundering altered the capability of the in vitro epidermal systems to absorb azinphos-methyl suspension. For paraoxon solution, however, new cotton glove fabric prevented absorption, and this protective effect, noted after 4 h of exposure, was lost when the fabric was laundered. Abrading the fabric did not cause a greater effect than laundering alone. These results suggest that the pesticide as well as its formulation may be factors of consideration when protective fabrics are chosen, and that, for cotton glove fabric, the protection against some OPs may best be provided before the fabric is laundered.

Absorption↗

PubMed1984

Prediction of the response to chemotherapy in acute leukemia by a short-term test in vitro.

To detect sensitivity or resistance of leukemic cells to chemotherapy prior to treatment, a short-term incubation method was employed. Blast cells from the peripheral blood or bone marrow of adult patients presenting with different forms of acute leukemia were analyzed for in vitro responsiveness to anticancer drugs in terms of suppression of 3H-uridine incorporation into cellular nucleic acids. The following agents were tested over a wide range of concentrations: Adriamycin, cytosine arabinoside, thioguanine, 6-mercaptopurine, prednisone, and 4-hydroperoxycyclophosphamide. Retrospectively, the in vitro data were compared to the clinical response of the patients to polychemotherapy. In the majority of the patients, in vitro cytotoxic effectiveness of Adriamycin (doxorubicin) and cytosine arabinoside reflected the in vivo situation. The levels of in vitro inhibition that could distinguish between drug-sensitive and drug-resistant diseases appeared to be 30% for Adriamycin and 20% for cytosine arabinoside. No correlation was found between the Adriamycin effect in vitro and the proliferative state (rate of 3H-thymidine incorporation) of the leukemic cell population. Serial in vitro sensitivity testing during the course of disease of various patients proved the ability of the test system to detect acquired resistance to chemotherapeutic agents. Therefore, the assay might serve as a reliable tool in the selection of effective chemotherapy in individual patients suffering from acute leukemia.

Acute Disease↗

PubMed2001

Expression of Cu/Zn and Mn superoxide dismutases during bovine embryo development: influence of in vitro culture.

Temporal pattern of expression of Cu/Zn and Mn superoxide dismutases (SODs) was investigated in bovine oocytes and embryos produced in vitro in two different culture conditions and in vivo after superovulation. SODs were examined at a transcriptional level in single oocytes and embryos by reverse transcriptase-polymerase chain reaction (RT-PCR) and, at a protein level, by Western blotting on pools of embryos. mRNA encoding Cu/Zn SOD were detected in in vitro bovine embryos throughout preattachment development as well as in in vivo derived morulae and blastocysts. Transcripts for Mn SOD gene were detected in most immature and in vitro matured oocytes as well as in some zygotes and 5- to 8-cell embryos while no transcript was found at the 9-to 16-cell stage in both culture conditions. In vitro embryonic expression of Mn SOD was detected earlier in the presence of serum. Half of the morulae showed the transcript if cultured with 5% serum while none without serum. At the blastocyst stage Mn SOD could be detected independently of culture conditions. For in vivo-derived embryos Mn SOD transcripts were detected both in morulae and blastocysts. Immunoblotting analyses revealed that Cu/Zn SOD and Mn SOD were also present at a protein level in in vitro-derived zygotes and blastocysts. Together these data demonstrate, for the first time, that Mn SOD is transcribed and that Cu/Zn and Mn SOD proteins are expressed in preimplantation bovine embryos. Finally, they suggest that Mn SOD transcription is altered by in vitro culture conditions.

Animals↗

PubMed1994

Radical solutions and cultural problems: could free oxygen radicals be responsible for the impaired development of preimplantation mammalian embryos in vitro?

A major obstacle to the study of mammalian development, and to the practical application of knowledge gained from it in the clinic during therapeutic in vitro fertilisation and embryo transfer (IVF-ET), is the propensity of embryos to become retarded or arrested during their culture in vitro. The precise developmental cell cycle in which embryos arrest or delay is characteristic for the species and coincides with the earliest period of embryonic gene expression. Much evidence reviewed here implicates free oxygen radicals (FORs) in the process of arrest. Thus, studies on the development of mouse preimplantation embryos in vitro have shown that (i) FORs are elevated in vitro, but not in vivo, at the time at which embryos become arrested or delayed, (ii) systems for removing reactive oxygen species to limit the formation of hydroxy radicals are present, although they have not yet been assessed quantitatively and may differ qualitatively from those in adult cells, (iii) metabolic and possibly genetic adaptations to oxidative damage are evident, (iv) published procedures for overcoming in vitro arrest are explicable in terms of FOR-mediated damage or responses and (v) the arrest or delay of most embryos in vitro can be reduced or prevented experimentally by addition of metal chelators to limit hydroxy radical formation and lipid hydroperoxidation.

Animals↗

PubMedDate not supplied

In vitro determination of tumour chemosensitivity in haematological malignancies.

A four-day tumour chemosensitivity assay of potential use in predicting tumour response to cytotoxic drugs has been developed for haematological cancers. The method comprised isolation of white cells from peripheral blood or bone marrow aspirates, drug exposure and incubation for 4 days. Drug-induced tumour cell kill was assessed by differential staining of live and dead cells such that the former could be morphologically identified. Tumour cell viability was subsequently calculated by reference to an internal standard of fixed duck red blood cells. Over 30 drugs have been tested in vitro, all of which have shown a dose response relationship in vitro and given a good scatter of sensitivities from patient to patient within the concentration ranges tested. In 27 cases where the in vitro chemosensitivity could be compared with the in vivo response, there were 7 true positive comparisons (sensitive in vitro and in vivo), 17 true negative comparisons (resistant both in vitro and in vivo) and 3 false positive comparisons (sensitive in vitro, resistant in vivo). A result was obtained in 38 of 50 samples received, comprising 16 of 18 chronic lymphocytic leukaemias, 11 of 20 acute lymphoblastic leukaemias, 5 of 5 acute myeloid leukaemias and 6 of 7 myelomas. The assay appears to show considerable promise as a tumour chemosensitivity test and warrants wider investigations.

Antineoplastic Agents↗

PubMed2006

In-patient versus in vitro degradation of P(L/DL)LA and PLGA.

To assess and compare degradation rates, poly(70L-lactide-co-30DL-lactide) [P(L/DL)LA] and poly(85L-lactide-co-15-glycolide) (PLGA) copolymer degradation were studied in patients (maxillofacial region) and in vitro. Five patients of a total of 20 with 85:15 PLGA osteosyntheses, and 15 of a total of 60 with 70:30 P(L/DL)LA osteosyntheses, (10 females and 10 males, 26-55 years; mean, 33 years) underwent explantations after 3, 6, 9, 12, 18, and 24 months. Identical resorbables immersed in physiological buffer solution at 37 degrees C were compared at identical intervals. P(L/DL)LA's initial average molecular weight of 45,000 decreased to 25,000 in patients (and to 21,000 in vitro) after 6 months, and to 8000 in patients after 18 months. PLGA's average molecular weight decreased from 44,600 to 22,000 after 3 months in patients and in vitro, and to 11,000 in patients and 1300 in vitro at 6 months. In-patient and in vitro glass-transition temperatures decreased from approximately 60 degrees C to 50 degrees C over 18 months. Crystallinity in explants was < or = 4% in P(L/DL)LA throughout testing, and 16% and 30% in PLGA at 6 and 12 months, respectively. Both copolymers decomposed reliably in patients: 85:15 PLGA within 12 months and 70:30 P(L/DL)LA within 24 months on average (p < 0.005), leaving only extremely small granules that powder upon finger touch. In vitro rates were significantly faster (p < 0.005). Crystallinity associated with foreign-body reactions was minor and did not inhibit decomposition.

Absorbable Implants↗

PubMed1983

Fertilizability of in vitro matured oocytes from golden hamsters.

The fertilizability of hamster oocytes matured in vitro was examined along with two factors potentially affecting nuclear maturation in culture. The four amino acids (isoleucine, methionine, phenylalanine, and glutamine) necessary for nuclear maturation of cumulus-free oocytes (Gwatkin and Haidri, '74) were not required if oocytes recovered on the morning of proestrus (day 4) were cultured with intact cumuli. Although follicular oocytes recovered on day 3 of the estrous cycle (late diestrus) had somewhat lower frequencies of maturation in vitro compared to those recovered on day 4 (76 vs. 95%, respectively), they still had a substantial frequency of spontaneous maturation. Follicular oocytes recovered on day 3 and matured in vitro were fertilized at frequencies equivalent to oviducal oocytes (80 vs. 82%, respectively) when incubation of oocytes with precapacitated sperm was continued for 6 h. Penetration of follicular oocytes was lower (37.4%) after only 4 h of sperm/egg incubation, indicating a delay in sperm penetration with follicular oocytes matured in vitro. Incubation for 4 h is sufficient time for penetration of 80% or more of oviducal oocytes. While 98% of penetrated oviducal oocytes were fertilized normally, only 2% of penetrated follicular oocytes were normal. The majority (85%) of follicular oocytes, unlike oviducal oocytes, were unable to cause decondensation of sperm nuclei after 6 h of sperm/egg incubation. Use of a highly defined system for in vitro fertilization of hamster gametes has provided rigorous proof that isolated cumulus-oocyte complexes do not undergo complete maturation in vitro.

Animals↗

PubMed1992

Developmental changes in the incidence of chromosome anomalies of bovine embryos fertilized in vitro.

In total, 196 two- to 32-cell bovine embryo and 104 blastocysts were obtained by the in vitro fertilization of follicular oocytes matured in vitro, and 15 blastocysts fertilized in vivo were used. Chromosomal anomalies in these embryos and the inner cell mass (ICM) separated immunologically were investigated. Chromosomal anomalies were observed in 12.1% (5/41) of 2-cell embryos, 20.0-36.4% of 4- to 16-cell embryos, 7.1% (1/14) of 17- to 32-cell embryos, 44.2% (15/34) of blastocysts, and 18.6% (13/70) of ICM cells derived from in vitro fertilization. These anomalies were mainly 3N and 4N at 2-cell stage, 1N and 1N/2N at 4- to 32-cell stages, and 2N/4N in blastocysts and in their ICM cells. Chromosomal anomalies of blastocysts from in vivo fertilization and their ICM were observed in 20.0% (1/5) of blastocysts and 33.3% (3/9) of ICM cells and these compositions were mainly 2N/4N. These results indicate that the abnormalities at early and blastocyst stages of embryos derived from in vitro fertilization were caused by abnormal fertilization in vitro and abnormal cleavage, respectively. Furthermore, a definite location of the chromosomal anomalies was observed in the trophectoderm of blastocysts derived from in vitro fertilization.

Animals↗

PubMed1982

In vitro skin evaporation and penetration characteristics of mosquito repellents.

An in vitro apparatus was used to study mosquito repellent evaporation and penetration characteristics with skin. The mosquito repellents 2-ethyl-1,3-hexanediol, N,N-diethyl-m-toluamide, N,N-diethyl-p-toluamide, 1-(butylsulfonyl)hexahydro-1H-azepine, and N,N'-dicyclohexamethyleneurea were studied. In vitro repellent duration, calculated from repellent evaporation rates, was compared to in vivo duration at the same dose (0.3 mg/cm2) to assess the validity of the model. In vitro durations for 2-ethyl-1,3-hexanediol, N,N-diethyl-m-toluamide, N,N-diethyl-p-toluamide, and N,N'-dicyclohexamethyleneurea correlated with in vivo durations (r2 = 0.94), although in vitro durations was longer than in vivo duration. 1-(Butylsulfonyl)hexahydro-1H-azepine, which had the longest in vivo duration, had an in vitro duration that exceeded the test period (12 hr). The 0-12-hr in vitro percutaneous penetration correlated with corresponding data available from in vivo studies.

Aedes↗

PubMed2002

Transgenic goats produced by DNA pronuclear microinjection of in vitro derived zygotes.

This study was undertaken to investigate various factors affecting the outcomes of in vitro fertilization (IVF) of oocytes retrieved by laparoscopic ovum pick-up (LOPU) technique from prepubertal and adult goats, as well as to evaluate the developmental competence of in vitro produced embryos. Oocyte-cumulus complexes recovered by LOPU from donors stimulated with gonadotrophins were matured in vitro. Fresh semen was used for IVF following various capacitation treatments. In vitro produced zygotes were either cultured to assess in vitro development or were transferred into recipients for full term development. The results indicated that successful IVF of the goat oocytes was affected by factors such as sperm capacitation treatment, oocyte quality, and abundance of cumulus cells on zona pellucida. Oocytes from both prepubertal and adult goats demonstrated similar full term developmental competence despite the fact that in vitro developmental rates were lower for prepubertal goats. The births of transgenic offspring demonstrated that the established LOPU-IVF technology combined with pronuclear microinjection can be successfully used to produce transgenic goats.

Animals↗