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Biomedical subjects

D Rath

Publications and source records attributed to D Rath.

At least 55 records · Page 3Linked to original sources
PubMed1997

[Reactions of non pregnant and and of cattle at different stages of pregnancy from the Holstein Friesian breed (HF) and from the old type German black and white breed (DSB) exposed to transport stress: variation and individual reaction patterns of hormone metabolic and hematologic variables].

Breeding animal transports between parts of our institute were used assessing effects of age of animals, of breed, of their functional state and of individual specificity of reaction patterns using peripheral venous blood samples. Increased mean values of Hb, Hk, body temperature, total protein, albumin, FT4, T4, FT3, T3, cortisol, adrenaline and decreased ones of body weight, Pa, Fe, noradrenaline were found after transport stress. Comparing age matched groups of DSB and HF breed higher concentrations of total protein, creatinine, lactate, cortisol, FT4, T3 and FT3 and lower ones of albumin, blood urea and Fe could be observed in animals of DSB breed before and after transport. Strong correlations existed on 14 variables within animals before and after transport stress meaning there could be individual specificity of some blood parameters.

Animals↗
PubMed1996

A randomised trial of two information packages distributed to new cancer patients before their initial appointment at a regional cancer centre.

The purpose of this study was to evaluate the extent to which a new patient information package (NPIP) or a mini version of the same package (mini-NPIP) reduces emotional distress and meets the informational needs of patients arriving at a tertiary cancer centre for the first time. A comprehensive package, NPIP, consisting of procedural information regarding cancer centre location, description of the health care team, treatment services, research, educational activities, accommodation and community services provided at the centre; and a condensed version of the same package, mini-NPIP, were developed. Consecutive patients with newly diagnosed breast, gynaecological, lung and prostate cancer, referred to the centre for the first time were prerandomised to receive NPIP, mini-NPIP or no information package. Patients randomised to NPIP or mini-NPIP were mailed the information package at least one week before their first appointment. On arrival at the centre, patients were administered the Brief Symptom Inventory (BSI) which measures psychological distress, and interviewed regarding preferences for information and acceptability of the information packages. Of 465 randomised patients, 161 were excluded post-randomisation and 304 completed the entire interview: 100 were randomised to the NPIP, 102 to the mini-NPIP and 102 to the control group. Emotional distress as measured by the BSI was similar for all groups (P = 0.98). Most patients preferred to receive the information (98%), receive it before the first appointment (84%) and by mail (79%). These preferences were more evident for those given the information packages. The majority of patients found the information packages easy to understand (88%) and useful (89%), and no differences were detected between packages. The cost of production and dissemination of NPIP was more than double the cost for mini-NPIP: $ 8.93 vs $ 3.98 (Canadian dollars) per patient. For patients presenting to a cancer centre for the first time, packages of procedural information do not appear to reduce psychological distress, but are preferred by patients. Given the cost of producing NPIP, mini-NPIP is the preferred approach.

Feasibility Studies↗
PubMed1996

Development of a culture medium (BECM-3) for porcine embryos: effects of bovine serum albumin and fetal bovine serum on embryo development.

Media are available that can deliver modest porcine embryonic development from a single-cell zygote to the blastocyst stage. However, few embryos develop to hatched blastocysts by Day 7 in vitro, indicating deficiencies in media that inhibit early embryonic development. A defined culture medium, Beltsville Embryo Culture Medium (BECM-3), was developed to support porcine zygote development to the blastocyst stage. When fetal bovine serum was added by late Day 5 (late morula/early blastocyst stage), 80% of total embryos cultured from Day 2 developed into hatched blastocysts by Day 8. There was also a significant increase in the mean cell number of blastocysts and hatched blastocysts when culture was performed in BECM-3-based media in the absence of BSA fraction V. These studies provide a chemically defined foundation for elucidating key developmental components of preimplantation pig embryos.

Animals↗
PubMed1996

Prospects for human reproductive medicine--applications of experimental techniques used in farm animal breeding.

Can assisted reproduction techniques performed in farm animal breeding be considered as future methods in human sterility treatment? The answer is no because, at the moment, the goals in human and animal reproduction are totally different. The methods that scientists are using or developing for farm animals are either without any therapeutic value for humans or they are forbidden by the German Embryo Protection Law. However, there are a few aspects which might be of value in the future of human assisted reproduction. The technique of in vitro maturation of immature follicles and oocytes, which is prerequisite in animal in vitro fertilization the female gamete pool after slaughter, might be used to obtain mature human oocytes from unstimulated cycles in women who do not respond to hormonal stimulation. Further more, it can be used in cancer patients to save their oocytes shortly before cancer treatment. Although still forbidden at the moment by the Embryo Protection Law, it is possible that the pre-implantation diagnosis of early embryos by the polymerase chain reaction technique after blastomere biopsy will become a valuable tool in preventing genome aberration in future.

Animals↗
PubMed1995

In vitro development to blastocysts of early porcine embryos produced in vivo or in vitro.

The objective of this study was to compare the development of porcine embryos from the 2- and 4-cell stages to the blastocyst stage after in vivo or in vitro fertilization and in vivo or in vitro culture. Early-stage embryos were collected either from superovulated gilts 36 h after the second mating or after in vitro fertilization (IVF) of in vivo-matured oocytes, both followed by in vitro culture to the blastocyst stage. Blastocysts collected from superovulated donors served as controls. In the first experiment, a total of 821 2- and 4-cell embryos derived from in vivo-fertilized oocytes was cultured either in medium NCSU 23, modified Whittens' medium or modified KRB for 5 d. Significantly (P < 0.05 and P < 0.001) more embryos overcame the 4-cell block and developed to the blastocyst stage in medium NCSU 23 than in the 2 other culture media. Hatching was only observed in medium NCSU 23. In the second experiment, embryos derived from in vivo-matured oocytes fertilized in vitro were cultured in medium NCSU 23. Of 1869 mature oocytes 781 (41.8%) cleaved within 48 h after in vitro fertilization. A total of 715 embryos was cultured to the morula and blastocyst stages, and 410 (57.3%) overcame the developmental block stage, with 358 embryos (50.1%) developing to the morula and blastocyst stages. None of the embryos hatched, and the number of nuclei was significantly (P < 0.05) lower compared with that of in vivo-fertilized embryos (18.9 +/- 9.8 vs 31.2 +/- 5.8). In the third experiment, 156 blastocysts derived from in vitro fertilization and 276 blastocysts derived from in vivo fertilization and in vitro culture were transferred into synchronized recipients, while 164 blastocysts were transferred immediately after collection into 6 recipients, resulting in a pregnancy rate of 83.3%, with 35 piglets (on average 7.0) born. From the in vitro-cultured embryos, 58.3% (7/12) of the recipients remained pregnant at Day 35 after transfer, but only 33.3% maintained pregnancy to term, and 14 piglets (on average 3.5) were born. In contrast, the transfer of embryos derived from in vitro-fertilized oocytes did not result in pregnancies. It is concluded that 1) NCSU 23 is superior to modified Whittens' medium and modified KRB and 2) blastocysts derived from in vitro fertilization have reduced viability as indicated by the lower number of nuclei and failure to induce pregnancy upon transfer into recipients.

Journal Article↗
PubMed1995

Collection of oocytes from cattle via follicular aspiration aided by ultrasound with or without gonadotropin pretreatment and in different reproductive stages.

Ultrasound-guided follicular aspiration was performed on 29 Holstein-Friesian cows/heifers twice weekly at 3- to 4-d intervals over a period of 2 consecutive estrous cycles (total 42 d). For visualization of the ovaries and guidance of the aspiration needle, a 6.5 MHz fingertip probe on a 62 cm probe carrier was inserted into the vagina. The disposable aspiration needle was connected to a permanent rinse tubing system, thus ensuring minimum death of oocytes in the aspiration processs. After penetration of the vaginal wall, the needle was inserted into a follicle of the rectally fixed ovary. Cumulus oocyte complexes (COC) were aspirated at a pressure of 100 mm Hg. In the first experiment, the effect of an additional gonadotropin treatment 4 d prior to aspiration was investigated in 8 lactating cows. Following FSH-treatment, the number of aspirated follicles was higher (P < 0.05) than in the nontreated animals (10.6 +/- 0.7 vs 8.9 +/- 0.5). The number of recovered COC (7.0 +/- 0.6 vs 5.8 +/- 0.5), the recovery rate (COC per aspirated follicle) (66.6% vs 65.4%), the percentage of viable COC (56.8% vs 52.1%), the cleavage rate upon in vitro maturation and in vitro fertilization (56.7% vs 59.8%) as well as the rate of morula/blastocyst formation (3.8% vs 2.9%) were similar in both groups. In the second experiment, follicles were aspirated in 4 lactating cows, 6 dry cows, 4 pregnant cows (first 35 d of pregnancy), and 4 heifers. The average number of aspirated follicles and recovered COC was higher (P < 0.05) in the first 2 groups (10.6 +/- 0.6 and 9.3 +/- 0.7 follicles; 7.2 +/- 0.5 and 6.9 +/- 0.7 oocytes) than in trie 2 other treatment groups (7.3 +/- 0.5 and 8.1 +/- 0.5 follicles; 5.0 +/- 0.4 and 5.7 +/- 0.5 oocytes). The percentage of viable COC was higher (P < 0.05; 68.3%) in lactating animals than in all the other groups (49.7, 52.5 and 57.4%, respectively). Similarly, upon in vitro fertilization, cleavage rate was higher (P < 0.05; 63.4%) in lactating cows than in the other groups (43.7, 50.5, 55.1%, respectively). A total of 21.5, 22.7, 11.9 and 13.5%, respectively, in the 4 groups of the in vitro fertilized oocytes reached the morula and blastocyst stages. After transfer of a total of 48 embryos 22 pregnancies (45.8%) were established as detected on Day 65. We conclude that 1) repeated aspiration of viable COC at short intervals is possible, 2) additional FSH-treatment does not increase oocyte yields, and 3) viable blastocysts can be produced from cattle at various reproductive phases irrespective of the reproductive phase.

Journal Article↗
PubMed1995

In vitro maturation of porcine oocytes in follicular fluid with subsequent effects on fertilization and embyo yield in vitro.

The objective of the present study was to test the ability of porcine follicular fluid (pFF) to improve maturation of porcine cumulus-oocyte-complexes (COC) in vitro and to observe subsequent effects on fertilization and development to late morula/blastocyst stages under in vitro conditions. The COC were incubated in Tissue Culture Medium (TCM) 199, supplemented with 1% fetal calf serum (FCS), 10% pFF collected from immature follicles (2 to 5 mm), with or without addition of 1microg/ml FSH. Control groups were matured in TCM 199 with or without FSH. Follicular aspirates were centrifuged (1700 x g, 5min.) and the supematants were stored at -20 degrees in 1.5-ml Eppendorff cups until used. On 7 experimental days a total of 3849 immature COC was aspirated from follicles ranging from 2 to 5 mm in diameter. A total of 1117 COC was selected for the experiments, and 239 COC were fixed and stained with 1.5% aceto-orcein after 48 h of in vitro maturation at 39 degrees C with 5% CO(2) in humidified air. Germinal vesicle breakdown (GVBD; 91.7%) and development to metaphase II (60.4%) were superior (P <or= 0.05) when 10% pFF and 1microg/ml FSH were present in the maturation medium. Another 878 COC were mechanically denuded and used for in vitro fertilization (IVF). Eighteen hours after IVF, 561 oocytes were fixed and stained with aceto orcein. No diffeences were found among groups at this time point. Another 317 embryos were continuously cultured to morula and blastocyst stages in medium NCSU 23. Cleavage rates differed significantly (P <or= 0.05) 48 h after IVF, and were superior (52.5% vs 32.5% vs 30.7%) when oocytes were matured in maturation medium supplemented with pFF and FSH as compared with maturation rates in TCM 199 without pFF and/or FSH. Once the embryos had cleaved, no further differences were observed in the development to morula and blastocyst stages. These data indicate that GVBD and nuclear matuation to metaphase II are enhanced by a combination of pFF and FSH and that cytoplasmatic maturation is more complete and better synchronized as indicated by the increased cleavage rates.

Journal Article↗
PubMed1994

Ovulation, fertilization and pronucleus development in superovulated gilts.

Estrus was synchronized in 45 gilts by ingestion of Zinc-Methallibur in the feed for 15 d. On Day 16 each gilts was treated with PMSG (1200 IU i.m.) followed in 72 h by hCG (500 IU i.m.). Gilts were inseminated 24 and 36 h after the onset of estrus followed by slaughter of groups (n = 4 or 5) at 40 h, 44 h, 48 h, 52 h, 56 h, 60 h and 64 h after hCG injection. Ovaries were evaluated macroscopically and oocytes/embryos were recovered by flushing the oviducts. The ovulation rate increased from 38% to 87% from 40 to 45 h and remained constant thereafter. At 40 h, 36% of oocytes were penetrated by a single spermatozoon. The rate of fertilization increased from 36% (40 h) to 59% (44 h), to 65% (48 h), to 73% (52 h), to 76% (56 h), 80% (60 h) and to 64% (64 h). At 40 h all fertilized ova contained a decondensed sperm head. After another 4 to 8 h early pronuclei were common, and 52 h after hCG treatment opposed pronuclei were predominant. The first cleavages were recorded 64 h after hCG injection.

Journal Article↗
PubMed1994

Successful gamete intrafallopian transfer (GIFT) in the porcine.

Gamete intrafallopian transfer (GIFT) was successfully established in the pig. In Experiment 1 (6 replicates) 234 oocytes (39 +/- 5.5 per recipient) plus spermatozoa (4000 to 8000 per oocyte) were transferred bilaterally into the oviducts of synchronized gilts, and embryos were recovered 48 h thereafter. The recovery rate was 50.4% and 50% of the recovered oocytes were fertilized. A total of 55 embryos was cultured in vitro in NCSU-medium for 48 h and 63.6% developed to morula or blastocyst stages. In Experiment 2 (5 replicates) 220 oocytes (44 +/- 4.9 per recipient) plus spermatozoa (4000 per oocyte) were transferred to 5 recipients which were allowed to go to term. Three gilts delivered 16 (n = 3, 5, 8) piglets. In Experiment 3 (5 replicates) 183 oocytes (36.6 +/- 1.2 per recipient) plus flow cytometry gender sorted spermatozoa (4000 per oocyte) were transferred to 5 recipients. The recovery rate was 47.8%, and 27.6% of the oocytes were fertilized. From all cleaved oocytes 45.8% developed to expanded blastocysts, with the number of blastomeres varying from 20 to 85 (38.3 +/- 22.5). These results indicate that the GIFT procedure can be used successfully in pigs, and can be a valuable tool for the study of gamete interaction as well as in the continued development of biotechnological procedures such as sex pre-determination.

Journal Article↗
PubMed1994

Differences in pronucleus formation and first cleavage following in vitro fertilization between pig oocytes matured in vivo and in vitro.

To elucidate the developmental differences occurring after in vitro fertilization (IVF) of pig oocytes matured either in vitro (n = 1934) or in vivo (n = 1128), the present experiment investigated the morphological changes from penetration to the two-cell stage. Oocytes were examined every 2-4 h from 2 to 32 h after in vitro insemination to study sperm penetration, male and female pronucleus formation, synkaryosis and first cleavage. The penetration rate was significantly higher (P < 0.05) for in vivo matured oocytes (69.8%) than for in vitro matured oocytes (35.0%). Penetration of spermatozoa into the ooplasm was first recorded 6 h (in vitro matured oocytes) and 4 h (in vivo matured oocytes) after addition of the spermatozoa to the oocytes. For both in vivo and in vitro matured oocytes, 2 h were required for sperm head decondensation. However, maximum sperm head decondensation occurred 2 h later in in vitro matured oocytes. Within 6 h, 41.7 +/- 5.6% of the in vivo matured oocytes had completed second meiotic division, whereas only 20.8 +/- 6.5% of the in vitro matured oocytes reached this developmental stage (P < 0.01). For in vitro matured oocytes, male pronucleus formation was retarded 2-4 h after onset of insemination and development of the female pronucleus was enhanced compared with in vivo matured oocytes. Synchronized opposing pronuclei were observed 14 h after insemination in in vitro matured oocytes and after 8 h in in vivo matured oocytes. Synkaryosis was first observed at 16 and 18 h in in vivo and in vitro matured oocytes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Transmural saturation transfer analysis of the creatine kinase system in the mammalian heart.

31P NMR spatial localization and saturation transfer techniques were combined to enable the transmural measurement of the forward creatine kinase (CK) rate (ATP:creatine N-phosphotransferase, EC 2.7.3.2.) in the in vivo canine myocardium. Five epicardial towards endocardial regions of the left ventricle (LV) were simultaneously examined using spatially localized voxels. Although intraleft ventricular CP/ATP ratios were constant, the pseudo first order rate constant (k') and the forward creatine kinase rate (Rf) displayed a 61% variation across the LV wall. Because CK levels and calculated [ADP], [CP] and pH are transmurally invariant in the normal left ventricle, the observed changes in the Rf could not be explained by changes in the absolute levels of these substrates and of creatine kinase. In addition, because myocardial oxygen consumption rates are known to be higher in the endocardium, these results imply that forward creatine kinase rates are not directly related to oxidative phosphorylation rates.

Adenosine Diphosphate↗
PubMed1992

Experiments to improve in vitro fertilization techniques for in vivo-matured porcine oocytes.

Three experiments were designed for the in vitro fertilization of in vivo matured oocytes following different sperm treatments: In experiment I, spermatozoa (sperm rich fraction) were capacitated in isolated and ligated uterine horns of estrous gilts (Method A) for at least 3.5 hours and 1x10(6) sperm/ml were exposed to mature oocytes. Of 1586 oocytes 509 (32.1%) developed to the two to eight cell stage. Cleavage stage embryos (n=187) were transferred into nine recipients; 81 embryos (43.3%) were recovered after 72 hours and 21 (26%) had developed to the morula or blastocyst stage. The average number of nuclei (whole mount staining) was 108.4 and indicated normal developmental capacity. Another 127 embryos were transferred into three recipients, two became pregnant and one gilt of them delivered two offspring after 116 days of gestation. In experiment II, two different procedures for the capacitation of spermatozoa using modified TCM 199 medium were compared with that of Method A. Semen was either adjusted to a concentration of 2x10(8) sperm/ml with modified TCM 199E (Method B), or after centrifugation the sperm pellet was diluted in a 1:1 ratio with the same extender (Method C). Motility, and hyperactivity were significantly different (P 0.05) among treatments and were best after uterine incubation (Method A). The percentage of head-to-head aggregation increased significantly (P 0.05) after incubation in the uterine horn, and is suspected to represent a spontaneous acrosome reaction. A total of 492 oocytes were fertilized with semen capacitated by the three methods however, maximal fertilization (34.7%; P 0.05) was obtained with Method B. In experiment III, Method B was repeated and in order to minimize the rate of polyspermy, a total of 298 oocytes were exposed to different sperm concentrations (8x10(3); 4x10(3); 2x10(3) per oocyte). The reduction of spermatozoa exposed to oocytes improved the fertilization rate significantly (P 0.05) from 37.2 to 54.5%.

Journal Article↗
PubMed1991

The accuracy of clinical findings and laparoscopy in pelvic inflammatory disease.

The accuracy of clinical diagnosis for pelvic inflammatory disease was determined in 95 women who presented with pelvic pain to primary care physicians and then were referred to gynecologists. Laparoscopy or laparotomy with endometrial biopsy and fimbrial minibiopsy revealed that prevalence of pelvic inflammatory was 46% (44/95) and positive and negative predictive values of gynecologists were 74% (23/31) and 67% (43/64) (p = 0.0002). If histopathologic diagnosis was the standard, clinical accuracies of the gynecologists were no better than chance (p = 0.43), suggesting an expectation bias for visual diagnosis. Laparoscopy had a sensitivity of 50% (12/24) and a specificity of 80% (40/50) for salpingitis if the standard was fimbrial histopathologic diagnosis (p = 0.01). These results support the routine use of laparoscopy, supplemented when negative by endometrial and fimbrial minibiopsy, to accurately diagnose pelvic inflammatory disease.

Cervix Uteri↗
PubMed1991

Scintigraphic measurement of regional gut transit in idiopathic constipation.

In this study, total gut transit and regional colonic transit in patients with idiopathic constipation were measured scintigraphically. Eight patients with severe constipation were studied, none of whom had evidence of abnormal function of the pelvic floor. 99mTc-radiolabeled Amberlite resin particles (average diameter, 1 mm; Sigma Chemical Co., St. Louis, MO) with a mixed meal were used to assess gastric emptying and small bowel transit; similar particles labeled with 111In were ingested in a coated capsule that dispersed in the ileocecal region. These were used to quantify colonic transit. Five healthy volunteers were also studied. Two patients showed delayed gastric emptying and two had slow small bowel transit. Seven of the eight patients had slow colonic transit. In five, delay affected the whole colon ("pancolonic inertia"); in two, transit in the ascending and transverse colon was normal, but solids moved through the left colon slowly. Mean colonic transit was also measured using radiopaque markers; this technique identified the patients with slow transit, as shown by measurements of overall colonic transit by simultaneous scintigraphy. However, estimated transit through the ascending and transverse colons was considerably shorter by the radiopaque marker technique. In conclusion, idiopathic constipation is characterized by either exaggerated reservoir functions of the ascending and transverse colons and/or impairment of propulsive function in the descending colon. Particle size may influence the result of regional colonic transit tests. Transit delays in other parts of the gut suggest that, in some patients, the condition may be a more generalized motor dysfunction.

Adult↗
PubMed1988

The presence of an additional fetal membrane and its function in the newborn guanaco (Lama quanaco e ).

The newborn guanaco (Lama quanaco e ) is covered by an extra fetal membrane. It shares this characteristic with the newborns of old-world camelids. This membrane, which is lubricated by the watery amnionic fluid typical for camelids, facilitates the delivery of the conceptus. the newborn gets rid of this membrane itself, since in camelids mothers do not lick their offsprings.

Journal Article↗
PubMed1987

[Practicality of ultrasound diagnosis in veterinary andrology].

It is possible to use the ultrasound technique in the veterinary andrology. Important results of that method may increase the evidence of the clinical examination of the male genital organs. To make sure that no harm may effect the sperm cells, ejaculates of bulls and boars were exposed to pulsed ultrasound (5 MHz/0,140 mW) up to 120 minutes. It was confirmed that this technique has no influence on sperm cell motility and morphology.

Animals↗