Abdominal radical trachelectomy (ART) performed during pregnancy: a case report.

PURPOSE OF INVESTIGATION: Cervical cancer is one of the most frequent malignant diseases diagnosed during pregnancy. Abdominal or vaginal radical trachelectomies are fertility-preserving alternatives to radical hysterectomy for young women with early-stage cervical cancer that can be performed during ongoing pregnancy. MATERIAL AND METHODS: The authors report a pregnancy complicated by cervical cancer treated by abdominal radical trachelectomy (ART) at 16-17 gestational weeks with preservation of the concurrent pregnancy. RESULTS: The pregnancy evolved normally and delivery occurred at 38-39 gestational weeks by elective caesarean section. CONCLUSIONS: Radical trachelectomy could be offered as an option for pregnant patients with early invasive cervical cancer. It may help women to avoid the triple losses of a desired pregnancy, fertility, and motherhood.

[Management of emergency cases, hospitalized in Romania in 2007]. / Date de management spitalicesc privind cazurile de urgenta din România in 2007.

UNLABELLED: We evaluated the main aspects of the management of emergency cases, hospitalized in Romania in 2007, by following the frequency of main diagnosis, their distribution by place and hospital and hospital indicators. MATERIAL AND METHOD: We collected our data from the Insurance Houses and National Institute of Statistics. RESULTS: Emergency cases represented in 2007 half of total cases hospitalized in Romania. Women are more frequent hospitalized then man or children. The Romanian South-East area had the highest frequency of emergency cases hospitalized. From all types of deceased cases, the emergency cases had the highest frequency. In emergency, the main category of diagnosis by frequency is "pregnancy, birth and recently given birth", much higher than non-emergency cases registered, also at the Obstretics-Gynecology and Emergency Hospitals. The medium period of days hospitalized for emergency cases was one week time. Most of the emergency cases are healed comparing to non-emergency ones. CONCLUSIONS: The main problem of hospitals in Romania is related to health services financing, costs for health care are still growing but also did the quality of health services and number of cases coming in emergency rooms.

Improving reproductive performance in lactating dairy cows by synchronizing ovulation or inducing oestrus.

Lactating crossbred Holstein-Friesian dairy cows (n = 331) were started on an Ovsynch regimen 68 +/- 8.2 days after calving; 200 micrograms GnRH intramuscularly (i.m.) on Days 0 and 9, and 35 mg prostaglandin F2 alpha i.m. on Day 7. Thirty-eight and 31 cows (11.5 and 9.4%, respectively) were in oestrus on Days 0 to 6 and 7 to 8, respectively, and inseminated, and the remainder were fixed-time inseminated (on Day 10). For these three groups, pregnancy rates (60-65 days after breeding) were 31.6, 38.7 and 34.0%, respectively (P = 0.82) and calving rates were 100, 100 and 89.9% (P = 0.23). In a preliminary trial, twelve lactating cows (45 to 60 days postpartum) with inactive ovaries were given 1500 IU eCG i.m.; 10 were in oestrus within 10 days after treatment (and inseminated) and eight of these were pregnant (30 days after breeding). The Ovsynch program resulted in acceptable reproductive performance in cyclic cows and eCG treatment has considerable promise for inducing oestrus in anoestrous cows.

Detection of calcium ionophore induced membrane changes in dog sperm as a simple method to predict the cryopreservability of dog semen.

The sensitivity of dog sperm cells for extracellular Ca(2+)/Ca(2+)-ionophore challenge was compared to the detrimental effects of an optimized freeze/thawing protocol. Three sperm-rich fractions of ejaculates from 9 dogs were obtained, and one aliquot of each ejaculate was washed in a modified Tyrode's medium (HBT containing 0.1 mM Ca(2+)), without (control sample) and with 2.5 microM Ca(2+)-ionophore (induced sample) and incubated for 60 min at 38 degrees C in humidified atmosphere. Another aliquot from the same semen fractions was diluted, washed in a Tris buffer, and packed into 0.5-ml straws with a Tris buffer containing 7.5 vol % glycerol. The samples were stored for 1 week in liquid nitrogen after a computer-driven three-step freeze protocol and subsequently thawed for 50 sec in a 37 degrees C water bath and reconstituted into HBT. The acrosome integrity was determined using fluorescein-conjugated peanut agglutinin (PNA-FITC) as an acrosomal marker, while the vitality of the sperm cells was simultaneously assessed with the membrane impermeable DNA supravital stain ethidium homodimer 1 (EthD-1) using fluorescence microscopy and flow cytometry. The motility of frozen/thawed sperm samples was evaluated by microscopic as well as computerized motility analyses. Remarkably, the percentage sperm cells that underwent acrosome reactions induced by Ca(2+)-ionophore correlated very positively (r = 0.93) with the amount of acrosome damage observed in cryopreserved sperm samples. Furthermore, the degree of cellular damage induced by Ca(2+)-ionophore treatment correlated very negatively (r = -0.99) with the relative amount of sperm cells that remained motile after cryopreservation. Such clear correlations between Ca(2+)-ionophore induced acrosome reaction and motility parameters for frozen/thawed dog sperm cells were not found, suggesting that the generation of acrosome leakage and sperm immotility are two independent detrimental processes occurring during cryopreservation. From these results it can be concluded that Ca(2+)-ionophore treatment followed by simultaneous determination PNA-FITC and EthD-1 staining can be used to predict the cryopreservability of ejaculates from individual dogs used as donors.

Comparative study of different methods for dog semen cryopreservation and testing under clinical conditions.

The extenders and freezing rates from three different freezing protocols were combined and compared to each other in order to study the post-thawing acrosome integrity and fertility of frozen dog sperm. A commercial bovine TRIS-base extender (TRILADYL) and two self-made canine semen extenders (Norwegian and Dutch) were combined with a conventional bovine and two canine freezing regimes, and acrosome integrity of frozen/thawed spermatozoa was assessed by fluorescein isothiocyanate conjugated peanut agglutinin staining (FITC-PNA). Differences between freezing/thawing protocols were reflected in the proportion of cells with acrosomal damage and not based on motility results. It was concluded that during dog semen cryopreservation extenders had less influence on the post-thawing sperm quality than did the freezing rates. The optimal extender/freezing rate combination (TRILADYL/Norwegian) was used in the clinical practice to evaluate the fertility of frozen sperm administered by intrauterine insemination using a surgical approach. The pregnancy rate was 57% (4/7), but the average litter size was low (2.8). This may have been due to the insufficient sperm numbers contained in an insemination dose and/or to the incorrect timing of artificial insemination (AI). The final conclusion is that the commercial bovine extender is useful for freezing dog semen, and the TRILADYL/Norwegian freezing protocol is recommended as the most advantageous combination for the freezing of canine semen in the clinical practice.

Analysis of serum and seminal plasma after feeding ochratoxin A with breeding boars.

The aim of the experiment was to investigate whether or not ochratoxin A (OA) can be detected in seminal plasma after feeding the toxin in five and 10 times of the human tolerable daily intake with breeding boars and how toxin profiles of serum and seminal plasma correspond to each other. In addition to that, the effect of the toxin challenge on motility and longevity of boar semen was also evaluated. OA from samples was analyzed by microplate ELISA. Percentage of progressive motility of spermatozoa was determined initially and after 24, 48, 96, 120 and 144 h of storage. OA appeared in serum and seminal plasma shortly after toxin application had started. Significant reduction of initial motility and impaired longevity was observed after toxin withdrawal. These findings suggest that OA might have the potential to affect sperm production and semen quality of boars, but further research is required to elucidate whether OA exerts direct effect on germinal epithelium or disturbs sperm cell maturation only.

Evaluation of sperm tail membrane integrity by light microscopy.

During routine evaluation of trypan blue-Giemsa stained semen smears, sperm cells can be found with unstained heads and with stained tails. It was hypothesized that these cells were immotile and should not be considered as live. Sperm motility was determined in isoosmotic, and presumably isotonic trypan blue-stained wet preparations. Bull, ram and boar semen smears were stained with hypoosmotic trypan blue-Giemsa to compare the relationship between the percentage of stained sperm tails and the percentage of sperm tails remaining straight under hypoosmotic conditions. Actively moving spermatozoa with unstained heads, but with stained tails were never observed in wet preparations. The correlation coefficient found between the percentage of sperm with stained tails and the percentage with straight tails was 0.81, 0.94 and 0.85 for bull, ram and boar spermatozoa, respectively. Results of this study show that sperm cells with an intact head membrane, but a stained and presumably membrane-damaged tail are not motile. Therefore these cells should be included in the dead category rather than alive in the usual live-dead studies with vital stains.

Morphologic, endocrine and thermographic measurements of testicles in comparison with semen characteristics in mature Holstein-Friesian breeding bulls.

Twenty Holstein-Friesian breeding bulls (62-79 months of age) were examined 3 times, at 30-day intervals. Scrotal thermograms for assessment of scrotal surface temperature (SST) and blood samples for plasma testosterone concentrations were taken just before and then 45 and 90 min, respectively, after treatment with GnRH (50 micrograms, Gonavet, i.m. per bull). Following GnRH treatment, there generally were significant increases in mean values of both top SST (range, -0.1 to 1.4 degrees C) and bottom SST (range, 0.3 to 1.8 degrees C). Scrotal circumference was highly repeatable but SST and video-measurements of scrotal dimensions were less repeatable, because apparently they were affected by ambient temperature. Plasma testosterone concentrations before GnRH treatment were more repeatable than those after GnRH treatment. Correlations between examinations of 0.67 to 0.81 and -0.14 to 0.47, respectively, but the converse was true for SST measurements. Semen was collected with an artificial vagina 3 times per week for 12 weeks starting 2 weeks before the first examination. The total number of spermatozoa per ejaculate was highly repeatable and the percentage of motile and live spermatozoa were relatively consistent. Separate regressions for each variable and for each examination were conducted for these 3 semen characteristics as dependent variables. For the number of spermatozoa per ejaculate and for the percentage of motile spermatozoa, significant independent variables were plasma testosterone concentrations and difference between top and bottom SST, respectively. The slopes of these equations were nearly all negative and the R2 was from 0.15 to 0.42. For prediction of the percentage of live spermatozoa, both SST gradient and plasma testosterone concentrations were significant independent variables. For these regressions, the slopes were negative and the regression coefficients were generally lower than for the other 2 dependent variables (range, 0.16 to 0.25). Treatment with GnRH and assessment of SST and plasma testosterone concentrations have some correlation with the semen production in the mature bull.

Endocrine and thermal responses to GnRH treatment and prediction of sperm output and viability in holstein-Friesian breeding bulls.

A study was conducted to determine changes in serum LH and testosterone concentrations and in scrotal surface temperature (SST; measured with infrared thermography) following GnRH treatment and to predict the number of spermatozoa collected and the proportion that were viable. Holstein-Friesian breeding bulls (n = 22, average age, 24.3 m.o.; range, 15 to 41 m.o.) were examined twice 30 d apart. Concurrently, semen was collected twice weekly with an artificial vagina. Treatment with GnRH (100 micrograms, i.m.) increased (P < 0.0001) serum LH and testosterone concentrations and increased (P < 0.0001) SST (range 0.6 to 1.1 degrees C; P < 0.05) at the top and bottom of the scrotum. In regression models to predict the total number of spermatozoa, significant independent variables included ultrasonic echotexture of the testes (negative slope), scrotal width (positive slope) and SST at the bottom of the scrotum 45 min after GnRH treatment (positive slope). In regression models to predict the percentage of live spermatozoa, ultrasonic echotexture was a significant independent variable (negative slope). Measurement of testicular ultrasonic echotexture and SST after GnRH treatment augmented measurement of testicular size for predicting the number and percentage of live spermatozoa.

Computer analysis of video and ultrasonographic images for evaluation of bull testes.

The objectives of this study were to determine relationships between scrotal size (SC; estimated from a video image) and testicular size, and between ultrasonographic echotexture of the testis and seminiferous tubule area in bulls. Video images of the scrotum of 49 Holstein-Friesian (H-F) bulls were recorded and digitized. Scrotal width and length were measured with custom software. After slaughter, scrotums (containing testes) were excised, SC and testicular height, width and volume were measured, and the testes were examined ultrasonographically. Correlations between SC and testicular width or volume (r = 0.86, P < 0.001 and r = 0.84, P < 0.001, respectively) were much higher than those between scrotal width and testicular width or volume (r = 0.23, P < 0.11 and r = 0.28, P < 0.06). Histological examination of the testes was performed in 31 of the bulls. Ultrasonographic echotexture of the testes (determined with custom software) was highly correlated (r = -0.5, P < 0.005) with seminiferous tubule area. Although SC was superior to video imaging for estimating testicular size, ultrasonographic imaging of the testes has considerable potential for the evaluation of testicular function in bulls.

Induction of acrosome reaction in dog sperm by calcium ionophore.

The sensitivity of the plasma membrane to calcium ionophore (A23187) challenge was studied in dog sperm using fluorescein lectin staining for the assessment of acrosomal status and viability. Second fraction ejaculates from 5 dogs were washed, resuspended in Ca(2+)-free (EDTA-treated), 50, 100, 500, 1000 and 2000 microM/l Ca(2+)-containing Sp-TALP medium and induced with 50, 250, 500, 1000, 2500 and 5000 nM/l calcium ionophore. Samples were collected from each aliquot after 30 and 60 min of induction to assess the percentage of acrosome reacted sperm cells (AR rate), viability and motility by fluorescein isothiocyanate conjugated peanut agglutinin (FITC-PNA) and ethidium-homodimer combined staining. On each slide, 200 sperm cells were assessed under epifluorescence microscope (x 1250) in a blind manner. The response to ionophore challenge (AR rate, viability, motility) varied with Ca2+ and ionophore concentration in the suspension. A significantly higher AR rate was detected in samples containing 100, 500, 1000 and 2000 microM/L Ca2+ (> 40%) than in that containing 50 microM/L. Acrosome reaction could not be successfully induced in the EDTA-treated sample and in any of the aliquots in which 50, 250 and 500 nM/L ionophore concentrations were used for induction. Motility decreased drastically in all of the treated samples and stopped in that sample where as significant AR rate could be detected. Viability remained high (> 75%) during the incubation and did not differ significantly in the treated and the control groups.

In vitro and in vivo motility studies of 99mTc HM-PAO labelled sperm cells.

The motility of 99mTc HM-PAO radiolabelled sperm cells, labelled as described previously (Balogh et al., 1992) was studied. The active migration of spermatozoa was demonstrated in capillary tubes containing bovine oestrous mucus, using an in vitro motility analyzer. Like nonlabelled sperm cells, the labelled spermatozoa covered a 4-5 cm distance in the capillary tubes during a 10-min run. In vitro motility testing of labelled spermatozoa from different animal species (cattle, rabbit, sheep, horse) did not reveal significant differences. The distribution of spermatozoa within the female genital tract was studied in a previously described animal model (rabbit) and in two new models (sheep, chicken). This method enables the determination and visualization of sperm distribution by a noninvasive technique. The results of in vivo motility tests pave the way for the introduction of a method of unprecedented specificity, which serves for studying the penetrability of the oviduct to spermatozoa.