Uterine Lavage Research Articles

The effect of spermatozoa and seminal plasma on leukocyte migration into the uterus of gilts.

Yorkshire x Landrace gilts were used to determine the effect of spermatozoa and seminal plasma on postbreeding uterine leukocyte influx. Estrus detection was performed with a boar at 12-h intervals following synchronization with 400 IU eCG and 200 IU of hCG. All gilts were AI once, 24 h after the detection of estrus following random assignment to a 2x2x3 factorial arrangement of treatments (sperm or sperm-free AI doses), AI dose medium (seminal plasma or PBS), and lavage time following AI. Gilts were treated with sperm (5x10(9) spermatozoa; SPZ; n = 30) or sperm-free (SF; n = 30) doses containing either 100 mL of seminal plasma (SP; n = 15/treatment) or PBS (n = 15/treatment). Uterine lavage was performed once on each gilt (n = 20/time) at one of three times after AI (6, 12, or 36 h) to determine the total number of uterine leukocytes. The leukocytes consisted predominately (92 to 99%) of polymorphonuclear neutrophilic granulocytes (PMN). There was an AI x medium interaction on uterine PMN numbers. The number of uterine PMN recovered from gilts inseminated with sperm suspended in PBS was greater than the number of PMN recovered from the uterine lumen of gilts inseminated with sperm in SP, SP alone, or PBS alone (P<.05). Furthermore, SP accelerated the rate of uterine clearance when suspended with sperm cells during the first 36 h following AI (P<.05). These results indicate that seminal plasma suppresses PMN migration into the uterus following breeding and enhances the rate of disappearance of uterine inflammation.

Influence of Hormone Replacement Therapy and Bacterial Inoculation on Proteins and Enzymes in Uterine Lavage Fluid of Ovariectomized Mares

Influence of Hormone Replacement Therapy and Bacterial Inoculation on Proteins and Enzymes in Uterine Lavage Fluid of Ovariectomized Mares

Minimally‐invasive early prenatal diagnosis using fluorescence in situ hybridization on samples from uterine lavage

A two-phase study was undertaken to examine the efficiency of using transcervical cells (TCCs) collected by uterine lavage and fluorescence in situ hybridization (FISH) for early prenatal diagnosis of fetal chromosome aneuploidy. Uterine lavage was performed in 50 women scheduled for elective termination of pregnancy (TOP, n=35) or chorionic villus sampling (CVS, n=15) between 6 and 11 weeks of gestation. TCCS were dissociated by trypsin and collagenase, and interphase FISH was carried out for chromosomes X, Y, 13/21, and 18. The phase I study comprised 36 women. The FISH results were compared with the cytogenetic analysis from long-term culture of villus samples collected at TOP or CVS. Among the 36 samples, 15 had a normal male karyotype and 21 had a normal female karyotype. FISH on TCCs correctly identified 13 out of the 15 pregnancies with a male fetus. In phase II, uterine lavage was performed on 14 women. The samples were first tested for the presence of trophoblasts with an anti-trophoblast antibody, GB25, by immunohistochemical staining. Among 12 GB25-positive samples, the FISH results corresponded to the fetal karyotype. One of the GB25-positive samples had five signals for the chromosome 13/21 probe. The cytogenetic analysis confirmed that the fetus had a karyotype of 47,XX,+21. In the GB25-negative samples, FISH failed to identify one male pregnancy. Follow-up was carried out on 13 ongoing pregnancies and no maternal or fetal complications were discovered. This study demonstrates that fetal chromosome numeration can be carried out using FISH on uterine lavage samples in early pregnancy. However, a specific fetal cell marker, such as specific anti-trophoblast antibody, is necessary to avoid a false-negative result. © 1997 John Wiley & Sons, Ltd.

Minimally-invasive early prenatal diagnosis using fluorescencein situ hybridization on samples from uterine lavage

A two-phase study was undertaken to examine the efficiency of using transcervical cells (TCCs) collected by uterine lavage and fluorescence in situ hybridization (FISH) for early prenatal diagnosis of fetal chromosome aneuploidy. Uterine lavage was performed in 50 women scheduled for elective termination of pregnancy (TOP, n = 35) or chorionic villus sampling (CVS, n = 15) between 6 and 11 weeks of gestation. TCCS were dissociated by trypsin and collagenase, and interphase FISH was carried out for chromosomes X, Y, 13/21, and 18. The phase I study comprised 36 women. The FISH results were compared with the cytogenetic analysis from long-term culture of villus samples collected at TOP or CVS. Among the 36 samples, 15 had a normal male karyotype and 21 had a normal female karyotype. FISH on TCCs correctly identified 13 out of the 15 pregnancies with a male fetus. In phase II, uterine lavage was performed on 14 women. The samples were first tested for the presence of trophoblasts with an anti-trophoblast antibody, GB25, by immunohistochemical staining. Among 12 GB25-positive samples, the FISH results corresponded to the fetal karyotype. One of the GB25-positive samples had five signals for the chromosome 13/21 probe. The cytogenetic analysis confirmed that the fetus had a karyotype of 47, XX, +21. In the GB25-negative samples, FISH failed to identify one male pregnancy. Follow-up was carried out on 13 ongoing pregnancies and no maternal or fetal complications were discovered. This study demonstrates that fetal chromosome numeration can be carried out using FISH on uterine lavage samples in early pregnancy. However, a specific fetal cell marker, such as specific anti-trophoblast antibody, is necessary to avoid a false-negative result.

Limb Defects as a Consequence of CVS or Uterine Lavage? A Need for Further Documentation of the Technique and Complications

Limb Defects as a Consequence of CVS or Uterine Lavage? A Need for Further Documentation of the Technique and Complications

Limb Defects as a Consequence of CVS or Uterine Lavage? A Need for Further Documentation of the Technique and Complications

Limb Defects as a Consequence of CVS or Uterine Lavage? A Need for Further Documentation of the Technique and Complications

P-224. Human seminal fluids and uterine lavages contain immunoreactive intact HCG that is not detectable by the Leydig cell bioassay

P-224. Human seminal fluids and uterine lavages contain immunoreactive intact HCG that is not detectable by the Leydig cell bioassay

Uterine lavage with saline in mares as treatment for endometritis

Uterine lavage with saline in mares as treatment for endometritis

SEVERE LIMB REDUCTION DEFECTS AFTER UTERINE LAVAGE AT 7–8 WEEKS' GESTATION

A 35-year-old multiparous Chinese woman underwent a uterine lavage procedure for prenatal sex determination at 7-8 weeks' gestation. The baby was delivered by elective Caesarean section because of breech presentation at 38 weeks' gestation. The child weighed 2425 g and had severe reduction defects of all four limbs. This case warrants advising prospective patients of a possible association and encouraging practitioners to mount a careful prenatal ultrasonography and postnatal follow-up of all pregnancies investigated by prenatal diagnostic uterine lavage.

Therapeutic considerations for mating-induced endometritis

A transient mating-induced endometritis is most likely physiological in the mare, serving to remove excess spermatozoa, seminal plasma, and contaminants from the uterus. However, the inflammatory reaction has to be resolved and inflammatory products cleared from the uterus prior to embryonal descent into the uterine lumen. Mares that fail to clear a semen-induced inflammation from the uterus develop a persistent mating-induced endometri t is which may result in infedi l i ty. l f susceptibi l i ty to persistent mating-induced endometri t is i suspected, the mare should be monitored by transrectal ultrasonography for intrauterine fluid accumulation at 6-1 2 hours intervals after breeding. Treatment should be aimed at assisting the uterus to physically clear contaminants and inflammatory products. Post-mating uterine lavage and/or the use of uterotonic drugs are recommended aI 6 12 hours after breeding in mares susceptible to persistent endometritis.

Effects of cervical dilation and intrauterine infusions on the timing of oviductal transport of equine embryos

Equine embryos spend 5 to 6 d in the oviduct before entering the uterus as expanded blastocysts, and cannot be consistently collected nonsurgically until Day 7. Technologies such as cryopreservation and embryo splitting, which are most successful with embryos at the morula or early blastocyst stage, have not been used in mares because equine morulae and early blastocysts are located in the oviduct and cannot be recovered nonsurgically. These experiments test the hypothesis that transport of equine embryos through the oviduct can be hastened by cervical dilation or by acute, sterile endometritis induced by intrauterine oyster glycogen treatment. Cervical dilation with or without intrauterine infusion of 0.5 ml PBS on Day 4 did not appear to hasten the transport of embryos into the uterus since Day 5 uterine embryo recovery rates were not higher (P > 0.1) for mares with cervical dilation or cervical dilation plus PBS infusion vs mares receiving no treatments (0 of 5 and 0 of 5 vs 0 of 10, respectively). Intrauterine infusions of 40 ml of 1% oyster glycogen or 40 ml of PBS on Day 3 did not appear to hasten the transport of embryos into the uterus since Day 5 uterine embryo recovery rates were not higher (P > 0.1) for oyster glycogen- or PBS-treated vs untreated mares (2 of 12 and 3 of 11 vs 0 of 10, respectively). Cervical and uterine treatments on Day 3 or Day 4 and uterine lavages on Day 5 decreased (P < 0.05) Days 11 to Day 15 pregnancy rates compared with that of untreated mares. Day 11 to Day 15 pregnancy rates were 1 of 5 for mares with Day 4 cervical dilation and Day 5 uterine lavage, 1 of 5 for mares with Day 4 PBS infusion and Day 5 uterine lavage, 2 of 12 for mares with Day 3 oyster glycogen infusion and Day 5 uterine lavage, and 3 of 11 for mares with Day 3 PBS infusion and Day 5 uterine lavage vs 7 of 10 for mares that received no treatment or lavage. Cervical and uterine manipulations on Day 3 or 4 and uterine lavage on Day 5 appeared to decrease pregnancy rates by Days 11 to 15. The results of these experiments do not support the hypothesis that cervical dilation or uterine infusion hasten oviductal transport, since neither cervical manipulation nor transcervical infusion of oyster glycogen or PBS into the uterus significantly hastened the rate of embryo transport into the uterus.

Methods for the transcervical collection of fetal cells during the first trimester of pregnancy

Before termination of pregnancy, four techniques for retrieving fetal cells transcervically were investigated: uterine lavage, endocervical lavage, cytobrush and mucus aspiration. The yield of fetal cells in these samples was studied and found to be somewhat better after uterine lavage. A preliminary assessment of the safety of mucus aspiration was carried out before transcervical chorionic villus sampling (CVS) in continuing pregnancies. No difference in outcome to a control group having only CVS was found.

Uterine cavity lavage: adding FISH to conventional cytogenetics for embryonic sexing and diagnosing common chromosomal aberrations.

This study was undertaken to examine the efficacy for early prenatal diagnosis of uterine cavity lavage at the level of the internal os and to assess the rate of maternal contamination. In phase I, uterine cavity lavage was performed in 38 women scheduled for pregnancy termination between 6 and 12 weeks. In addition to short- and long-term cultures, one-colour FISH (fluorescence in situ hybridization) with Y and X probes was used for fetal sexing. Two-colour FISH was used in all known male fetuses for the assessment of maternal contamination. In phase II, lavage was performed on 16 women. Fetal sex was diagnosed with direct labelled X and Y probes and common numerical chromosomal aberration was attempted with 18 and 21 direct labelled probes. Fetal sexing was successful in all cases in phases I and II. Out of 34 patients in which tissue was obtained, only FISH was done in six. Long-term cell cultures were successful in the other 28 cases, but complete karyotyping in 19 (56 per cent). No chromosomal aberration was found with the direct labelled probes 18 and 21 in FISH. Maternal contamination was assessed to be 5-10 per cent. This simple and easy-to-master technique is very effective in obtaining fetal cells early in pregnancy for genetic diagnosis, especially by FISH. However, the safety of the procedure must be tested in ongoing pregnancies.

Influência da lavagem uterina nos índices de concepção pós-parto em éguas puro-sangue de corrida

Avaliou-se o efeito da lavagem uterina pós-parto com solução de NaCl a 0,9% ou iodo-povidine a 0,4% em éguas Puro-Sangue de Corrida (PSC), em relação aos índices de concepção pós-parto (p.p.). Foram comparadas as taxas de concepção de 27 éguas, sem qualquer manipulação uterina (Controle), 57 éguas com solução de NaCl (I), 36 éguas com iodopovidine (II) e 15 éguas apresentando retenção de placenta, que sofreram lavagem uterina com as duas soluções. Concluiu-se que as lavagens uterinas p.p., independentemente da substância utilizada, influenciaram negativamente a involução e a preparação do útero para a concepção, só estando indicadas em éguas com retenção de placenta, desde que aplicadas até o 3º. ou 4º. dia p.p.

Oestrogen synthesis by the peri-implantation conceptus of the one-humped camel (Camelus dromedarius)

The ability of the embryonic tissues of the dromedary camel (Camelus dromedarius) to synthesize oestrogens in vitro was studied in 15 conceptuses recovered by non-surgical uterine lavage between 10 and 33 days after ovulation. Definitive evidence for strong aromatase activity with synthesis of considerable quantities of oestrogens was obtained at all stages when conceptus tissues were incubated with [3H]androstenedione. A high proportion of the oestrogens was in the form of oestradiol, which contrasts to the higher ratio of oestrone:oestradiol in the oestrogens synthesized by embryonic tissues of horses and pigs. Biopsies of endometrial tissue recovered from pregnant and nonpregnant camels showed great ability to conjugate both oestradiol and oestrone when incubated with tritium-labelled forms of these two hormones.

Immunoglobulin (IgG and IgA) and complement (C3) concentrations in uterine secretion following an intrauterine challenge of Streptococcus Zooepidemicus in mares susceptible to versus resistant to chronic uterine infection1

The validity of measuring concentrations of immunoglobulins in undiluted uterine secretions was established. The concentrations of IgG, IgA, and cleavage factor C3 of the complement system in uterine secretions were compared in mares with different resistance to chronic uterine infection (CUI). The uteri of mares susceptible (n = 6) and resistant to CUI (n = 5) were inoculated with 5 x 10(6) Streptococcus zooepidemicus when the mares were in estrus. Uterine secretions were sampled, and sampling was immediately followed by a uterine lavage at 5 and 24 h after bacterial inoculation. During a subsequent estrus, bacterial inoculation of the uterus was repeated, and samples were taken from the mares 12 and 36 h later. The fraction of uterine secretion in each uterine washing was determined by dividing the protein concentration of the undiluted uterine secretion by the protein concentration of the corresponding uterine washing. There was a significant correlation between measured concentrations of immunoglobulins in uterine secretions and calculated concentrations in the uterine washings (p < 0.05). Concentrations of IgG and C3 in uterine secretions declined similarly in both susceptible and resistant mares during the first 24 h after bacterial inoculation (p < 0.04). In contrast to the susceptible group, which showed a continuous decline at 36 h, resistant mares demonstrated an increased concentration of both IgG and C3 at this time. Concentrations of IgA did not differ between susceptible and resistant mares. It was concluded that the described method of sampling uterine secretions was useful for analyzing IgG and IgA.(ABSTRACT TRUNCATED AT 250 WORDS)

Progesterone and estradiol in biodegradable microspheres for control of estrus and ovulation in mares

Progesterone and estradiol 17-β in poly (DL-lactide) microspheres were used to control estrus and ovulation in mares after luteolysis was induced by prostaglandin F 2∞. Mares were given a single intramuscular injection of biodegradable poly (DL-lactide) microspheres, 1 day following prostaglandin treatment, containing no hormones (control), 0.625 g progesterone and 50 mg estradiol (low dose), 1.25 g progesterone and 100 mg estradiol (medium dose), or 1.875 g progesterone and 150 mg estradiol (high dose; n=15 mares per group). Mares treated with the low dose had significantly longer intervals (P<0.05) to estrus and ovulation than the control mares; however, low dose mares had shorter intervals (P<0.05) to estrus than high dose mares and shorter intervals to ovulation than medium and high dose mares. Regression analysis indicated that the medium dose was sufficient for maximizing interval to ovulation while the high dose maximized interval to estrus. All groups of mares exhibited similar (P>0.05) post-treatment estrus lengths. A clinical response scoring system based on synchrony of both estrus and ovulation within a treatment group was also used to measure the effectiveness of treatments on control of estrus and ovulation. Clinical response scores did not differ (P>0.05) among treatment groups. Mares were randomly assigned for insemination at the beginning of the first post-treatment estrus. Rates for embryo recovery performed by uterine lavage 7 days post-ovulation did not differ (P>0.05) among groups. Concentrations of serum progesterone increased in mares receiving progesterone and estradiol microspheres. At 10 to 14 days post-injection of microspheres, progesterone concentrations were higher (P<0.05) and remained above 1 ng/ml in the mares receiving the high dose. Progesterone concentrations were also higher (P<0.05) on Days −3 to −1 (Day 0 = day of post-treatment ovulation) in mares receiving the high dose when compared to control mares. Gonadotropin concentrations were suppressed (P<0.05) in the medium and high dose groups.

Trophectoderm microbiopsy in murine blastocysts: Comparison of four methods

This study compares four trophectoderm microbiopsy techniques for removal of blastomeres from murine blastocysts: (1) aspiration, trophectoderm pipetted through the zona; (2) incision, trophectoderm excised with a microrazor; (3) slit/excision, the zona slit and herniating trophectoderm excised; and (4) hatch/excision, trophectoderm cells excised after spontaneous hatching. Murine blastocysts were comparatively biopsied using one of four methods and contrasted to zona slit and nonmicromanipulated controls. Operative cellular injury was assessed by uptake of fluorescein isothiocyanate (FITC). Postoperative embryonic viability was assessed by blastocoele reexpansion and hatching inner cell mass development and trophectoderm plating. All techniques yielded cells available for genetic analysis. The slit/excision technique and hatch/excision techniques exhibited lower operative injury and the higher postoperative viability than aspiration or incision. The slit/excision and the hatch/excision techniques, though requiring two operative steps, appear to be the least damaging of the four methods. Therefore, they should be most applicable to human blastocysts obtained either by extended culture in vitro or by uterine lavage.

Assessment of the Reproductive System of the Female Ruminant

In summary, a thorough physical examination of the reproductive tract includes a complete history, general physical examination, and evaluation of the external and internal genitalia. Ancillary tests that can be applied to this system include uterine culture, biopsy, cytology, and lavage.

Videoendoscopic evaluation of the mare's uterus: III. Findings in the pregnant mare.

During a 5-year period 65 Pony and 20 Thoroughbred pregnant mares were subjected to videoendoscopic hysteroscopy from 10 to 266 days of gestation. The aims of these examinations were to 1) observe foetal and placental development in vivo (60 Pony and 10 Thoroughbred mares); 2) eliminate one of unicornuate twin conceptuses (9 Thoroughbred mares); 3) recover embryonic foetal and placental tissues non-surgically for experimental purposes (47 Pony mares); 4) induce focal separation of the placenta in late gestation as an experimental model of placentitis (5 pony mares and 1 Thoroughbred mare). It was possible to view the embryo and all constituent extra-embryonic membranes of the conceptus between Days 10 and 87 of gestation without having to perforate the allantochorion. This allowed study of physiological processes such as the coordinated uterine contractions responsible for conceptus motility between Days 7 and 17, active foetal movements, which began as early as Day 34, and invasion of chorionic girdle cells into the endometrium, which occurred between Days 34 and 38. From Day 90, vision of the foetus was reduced or prevented by the increased thickness of the allantochorion. Transendoscopic recovery of the conceptus was successful in all of 10 mares under 30 days of gestation, whereas only 10 of 18 attempts between Days 30 and 45 produced the conceptus without resorting to uterine lavage after initial rupture of the allantochorion. All 9 attempts to eliminate one of unilateral twin conceptuses were unsuccessful and the technique was abandoned as clinically unsuitable. Nevertheless, 9 of 22 (41%) single conceptuses remained viable after one or more hysteroscopic examinations.(ABSTRACT TRUNCATED AT 250 WORDS)