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Biomedical subjects

M Gilles

Publications and source records attributed to M Gilles.

18 recordsLinked to original sources
PubMed2006

The effect of nitric oxide inhibition and temporal expression patterns of the mRNA and protein products of nitric oxide synthase genes during in vitro development of bovine pre-implantation embryos.

This study was conducted to determine the effect of Nitric oxide (NO) inhibition in bovine in vitro development and expression analysis of the three Nitric oxide synthase (NOS) isoforms: endothelial (eNOS), neuronal (nNOS) and inducible (iNOS), mRNA and protein in bovine oocytes and embryos. Selective inhibitor of NOS, N-omega-nitro-l-arginine methyl ester (l-NAME) was applied at different doses (0, 0.1, 1 and 10 mm) in maturation (experiment 1A), culture medium (experiment 1B) and in both maturation and culture media (experiment 1C). No significant differences were observed in cleavage and blastocyst rates when oocytes were matured in the presence of l-NAME as long as the inhibitor was omitted during fertilization and culture. However, significantly lower blastocyst rates were observed when l-NAME was present at higher level (10 mm) in culture medium alone and in both maturation and culture media. In experiment 2, mRNA isolated from triplicate pools of oocytes and embryos (n = 15-20) was subjected to quantitative real time reverse transcription polymerase chain reaction to investigate the expression of eNOS, iNOS and nNOS mRNA in normal IVP bovine oocytes and embryos. While eNOS and iNOS transcripts were detected at higher level in oocytes (immature and mature), two-cell and four-cell stage embryos, the nNOS was detected only in immature oocyte, two-cell and morula stages. In experiment 3, eNOS and iNOS protein expression analysis was performed in IVP oocytes and embryos and both proteins were detected in the cytoplasm and the nuclei (weak) of oocytes and embryos. These data provide the first evidence for the role of NO production and the presence of mRNA and protein products of NOS isoforms during bovine embryogenesis.

Animals↗
PubMed2006

Transcript profiles of some developmentally important genes detected in bovine oocytes and in vitro-produced blastocysts using RNA amplification and cDNA microarrays.

To study the mRNA transcript profiles of some potential candidate developmental genes during bovine oocyte and blastocyst stages, RNA amplification procedures, cDNA microarray of 82 target genes spotted onto glass slide and real-time polymerase chain reaction (PCR) were used. Messenger RNAs were isolated from in vitro-produced bovine matured oocytes and blastocysts. Using equal amounts of input mRNAs but different cycles of amplifications, cDNAs were produced and served as template for RNA amplification by the in vitro transcriptions. After amplification, the RNA yields transcribed from cDNAs of different cycles were evaluated both by hybridization on the cDNA microarrays and by using real-time PCR techniques. The analyses indicated best results from lower amplification cycle templates with consistent signals at hybridization. Generally, the RNA yield was directly proportional to the amplification cycle but inversely related with signal consistency at repeated hybridizations. Using the protocols established, equal amounts of amplified RNA from matured oocytes and blastocysts were hybridized to the array. Analyses of replicated hybridizations indicated that 35 transcripts were differentially expressed. Most of these were not described in previous bovine embryo studies. Independent analyses of 23 transcripts with real-time PCR and unamplified RNA confirmed the results of 22 genes. Moreover, the functional analyses showed various roles related to development. Hence, it is possible to conclude that the genes identified here are potential candidates for characterizing developmental competence, and that the methods established can be used for large-scale gene expression analysis with more comprehensive arrays.

Animals↗
PubMed2005

Paroxetine serum concentrations in depressed patients and response to treatment.

INTRODUCTION: There is no established relationship between the serum concentration of selective serotonin reuptake inhibitors (SSRIs) and clinical response in depressed patients. METHODS: We analyzed paroxetine concentrations in serum of 46 depressed patients during treatment with a fixed dosage of 40 mg paroxetine. RESULTS: After 5 weeks 29 patients responded to treatment, while 17 did not. Analysis of variance with repeated measures (ANOVA-rm) revealed a significant effect of "response" with responders having lower serum concentrations throughout the treatment period, when compared to non-responders. After 2, 3, and 4 weeks of treatment, we could define an upper threshold of paroxetine serum concentrations (week 1 : 22.7 ng/mL; week 2 : 43 ng/mL; week 3 : 53.4 ng/mL; week 4 : 39.1 ng/mL) above which response to treatment was unlikely. CONCLUSION: We conclude that -- in contrast to other pharmacological approaches -- high rather than low drug serum concentrations may be associated with non-response in paroxetine treatment of depressed patients.

Adult↗
PubMed2005

Expression of retinoid X receptor transcripts and their significance for developmental competence in in vitro-produced pre-implantation-stage bovine embryos.

Retinoid X receptors are frequently implicated in their role in embryonic development. However, there is no report regarding their expression in embryos with different qualities. In this study, expression pattern and levels of retinoid X receptor transcripts in different qualities and stages of in vitro-produced pre-implantation bovine embryos were examined using real-time polymerase chain reaction (PCR) techniques. Four independent cDNA pools per sample were prepared from mRNAs isolated from different developmental-stage embryos, after classifying them to be of good and bad quality. Primers were also designed to amplify products with PCR. Based on analysis, except retinoid X receptor gamma (RXRG) transcripts, the remaining two types (alpha and beta forms) were expressed in all samples with significantly higher (p < or = 0.05) levels until the eight-cell stage compared with the stages thereafter. However, the transcript levels for bad-quality embryos were consistently lower in all cases in comparison with good-quality embryos. RXRG was identified in 39% of the samples, but all in the pre-embryonic genome activation development stage. Therefore, strong expression patterns of these transcripts in earlier stages indicates their possible role during the maternal phase of embryo development, and the variations of copy number in embryos with different qualities substantiates their possible candidacy as potential quality markers. Moreover, identifying the transcript variations in different qualities and expression of RXRG at these stages is a novel input to the pre-implantation-stage knowledge.

Animals↗
PubMed2004

Intrafallopian transfer of gametes and early stage embryos for in vivo culture in cattle.

It may be possible to avoid inadequate in vitro culture conditions by incubating gametes or embryos in the oviducts for a short time. Ideally, an optimized procedure should be devised, combining in vitro and in vivo systems, in order to achieve synchronization in cattle. We transferred gametes as well as embryos in various stages of development and placed them into the oviducts. Embryos were recovered on Day 7 by flushing of oviducts and uterine horns. Blastocyst rates were determined on Day 7 and on Day 8. Experimental designs included transfer of in vitro matured cumulus oocyte complexes into previously inseminated heifers (COCs group), transfer of in vitro matured COCs simultaneously with capacitated spermatozoa (GIFTs group), transfer of four to eight cell stage embryos developed in vitro after IVM/IVF (Cleaved Stages group) and a group of solely in vitro produced embryos (IVP control group). Our results indicate that in vivo culture of IVM/IVF embryos in the homologous bovine oviduct has a positive influence on subsequent pre-implantation development. In addition, we have evidence that in vitro maturation and in vivo fertilization cannot be synchronized.

Animals↗
PubMed2004

Open-label non-randomized versus double-blind randomized antidepressive treatment: what are the advantages of clinical decision over randomization?

OBJECTIVE: To study, whether and how the results from open and double-blind randomized trials on antidepressants differ. METHODS: Seventy-one patients were included in a study comparing open, non-randomized, standardized treatment with paroxetine (PAROX) and amitriptyline (AMI) after a minimum of six drug-free days (OPEN). A second group of 56 patients received the same treatment under blind-randomized conditions (BLIND-RANDOM). The course of psychopathology as assessed by the Hamilton Depression Rating Scale was compared using repeated measurements ANOVA-(rm). RESULTS: While the rate of adverse events was higher in the BLIND-RANDOM compared to the OPEN condition, completer-analyses revealed no differences in psychopathological outcome. CONCLUSIONS: With similar clinical outcome BLIND-RANDOM trials of antidepressants may expose depressed patients to an increased risk of adverse events, when compared to OPEN conditions. However, the clinical outcome in study completers did not differ between the BLIND-RANDOM and the OPEN condition. Thus, the psychiatrist's choice may have impact on adverse events rather than on clinical outcome of antidepressant treatment.

Adult↗
PubMed2004

A comparative expression analysis of gene transcripts in post-fertilization developmental stages of bovine embryos produced in vitro or in vivo.

This study was carried out to examine the temporal variation in the relative abundance of transcripts during the post-fertilization stages of bovine embryos derived from in vitro or in vivo culture. For this purpose, cumulus-oocyte complexes obtained from ovaries from slaughterhouses were matured, fertilized and cultured in vitro. The in vitro culture was carried out using CR1 medium. In vivo embryos were derived from superovulated and artificially inseminated Simmental heifers and cows. Embryos were recovered from both systems approximately at day 1 (two-cell), day 2 (four-cell), day 3 (eight-cell), day 4 (16-cell), day 5 (morula) and day 7 (blastocyst) after insemination. Relative abundance of target transcripts was performed using real-time PCR. The transcripts studied were the nucleosome assembly protein, mRNA for alivin 1, Pleckstrin homology Sec7 coiled domain, polyadenylate binding protein, NADH dehydrogenase subunit 2, high-mobility group transcription factor, cytokine-like nuclear factor, NY-REN-58 antigen mRNA, mRNA for KIAA1764 and one novel transcript. These transcripts were derived from our previous study conducted using mRNA differential display reverse transcription-polymerase chain reaction (DDRT-PCR) between in vitro-produced bovine eight-, 16-cell, morula and blastocyst-stage embryos. The results of the present study have demonstrated the temporal variation in the relative abundance of these transcripts between in vitro and in vivo culture systems. Such variation in the relative abundance of transcripts during the post-fertilization developmental stages of in vitro and in vivo embryos may reflect the effect of the in vitro culture condition on the transcriptional activity thereby on the developmental competence of the resulting embryos.

Animals↗
PubMed2003

Mapping of ESTs derived from pre-implantation stage cattle embryos to allocate 16 new additions to the ordered comparative map of cattle and human.

Twenty expressed sequence tags (ESTs) derived from cDNA libraries of different developmental stages of embryos were mapped using a whole genome bovine hamster radiation hybrid panel. These include 14 markers representing genes, most of which have not so far been mapped in cattle, with another three being novel in both cattle and human. The markers were placed on specific chromosomes with high LOD scores and except two all localizations fit the current human and cattle comparative map. The assignment of these genes further enriches the cattle genome map and also contributes to the international effort of generating comparative maps.

Animals↗
PubMed2001

Morphological and functional characteristics of the dominant follicle and corpus luteum in cattle and their influence on ovarian function.

Predicting the functional activity of a dominant follicle (DF) and corpus luteum (CL) might be important before starting a superovulation regime or a synchronization program. The DF and CL were characterized morphologically by using ultrasonography and were characterized functionally by estimating the estradiol-17beta/progesterone (E2/P4) ratio. Their influence on ovarian function was estimated through their ability to ovulate at different stages of development in response to PGF2alpha-application. A total of 47 Holstein Friesian (35 cows and 12 heifers) were used in two experiments. In Experiment 1, 25 animals were examined by daily transrectal palpation and ultrasonography to follow the morphological development of the DF. The status of the DF was categorized into 3 groups (A1, B1, C1). The A1 group (n=7) contained animals with DF in the growing phase or in early static growth phase for less than 3 days. Group B1 (n=13) included animals with DF in static growth phase for 3 to 4 days, while Group C1 (n=5) comprised animals with DF keeping a plateau for more than 4 days or animals with DF in the regression phase. The DF were aspirated transvaginally and the follicular fluid (FF) was analyzed for E2 and P4. In Experiment 2, 22 animals were included. As in Experiment 1, the animals were classified into three groups (A2, n=10; B2, n=5; C2, n=7). They were treated by a single dose of PGF2alpha (25 mg, i.m.) between Days 8 and 12 of the cycle. Results showed that luteolyses occurred in all animals. The DF, which were in growing or in early static growth phase < 3 days were always E2-dominant (E2 > P4) and ovulated after PGF2alpha-application in 6/8 of cases and persisted in 2 (Group A2). The DF persisting > 4 days or that had been in regression were always P4-dominant. This type of DF regressed after PGF2alpha-application (Group C2). The DF in early static growth phase for 3 to 4 days in 5/13 cases were E2-dominant and in 8/13 cases were P4-dominant. This type of DF ovulated in 3/5 cases and regressed in 2/5 cases after PGF2alpha-application (Group B2). These results suggest that the DF is morphologically and functionally defined as long as the DF is in the growing or early static growth phase (A1, A2) for at least 2 days or if the DF is in regression (C1, C2). However, when the DF is in the static growth phase for 3 or 4 days (B1, B2), their morphological and functional characteristics are different. The CL controlls ovulation in the A and C groups and plays an abettor's roll in the B-group.

Animals↗
PubMed2001

Increased platelet aggregability in major depression?

There is compelling evidence that depression constitutes an independent risk factor for cardiovascular morbidity and mortality. As exaggerated platelet reactivity is associated with an increased risk of intra-arterial thrombus formation, we studied platelet aggregability in patients with major depression both before and after 5 weeks of anti-depressant therapy as well as in healthy control subjects. Twenty-two depressed patients and 24 healthy control subjects participated in the study. Washed and rediluted platelets were stimulated with the agonists collagen and thrombin in three concentration steps. Depression was associated with a higher aggregability after stimulation with thrombin in the intermediate concentration and with collagen at the low concentration, with ceiling effects for the other concentrations. After 5 weeks of anti-depressant therapy, aggregability was somewhat less exaggerated, although this effect did not reach statistical significance. We thus conclude that major depression is associated with increased platelet aggregability, which seems to persist even under a marked improvement in depressive symptomatology. This effect may contribute to the increased cardiovascular morbidity in depressed patients.

Adult↗
PubMed2000

The 'three in one' block as locoregional analgesia in an emergency department.

The 'three in one' block is a well-known locoregional technique for per- and post-operative anaesthesia. This 'three in one' block was investigated in an emergency department with three combinations of local anaesthetics as a locoregional way of pain relief for patients with fractures of the upper femur. A double-blind prospective study was conducted with 61 consecutive patients. A good regression of the linear visual analogue scale (LVAS), good haemodynamic stability and great patient satisfaction were found for all groups of patients at different times after installation of the block. Side effects were few and are avoidable by reducing the amount of local anaesthetic given to elderly patients with a low bodyweight. The technique failed in only four patients due to technical reasons. It was concluded that the 'three in one' block is a fast, safe, reliable and easy technique for pain relief in an emergency department. Good analgesia was achieved for patients with fractures of the upper femur with no need to change the position of the patient.

Aged↗
PubMed1999

Lateral balance organisation in human stance in response to a random or predictable perturbation.

The effect of the predictability of perturbation to standing balance was evaluated in terms of the muscle activity and response dynamics of five subjects exposed to horizontal forces at the pelvis producing sideways or forward sway. Rapid (EMG onset latencies of 70-80 ms recorded from the left gluteus medius and gastrocnemius) and qualitatively different patterns of response were produced by forward pushes and pushes to either side. However, the EMG response to left push was constant in pattern and timing, whether the push direction was constant and, therefore, predictable over a block of trials or whether the left push trials were interleaved randomly with right push or forward push trials. Moreover, there were no systematic effects of perturbation direction uncertainty on the latency and rate of increase of ground reaction forces. We conclude that prior information does not speed postural responses that differ quantitatively according to the direction of perturbation to balance.

Adult↗
PubMed1999

Integration of the hand in postural reactions to sustained sideways force at the pelvis.

In order to investigate the potential hand contribution to sideways balance, sideways pushes to the right, which subjects resisted using either the lower limbs ("hip only") or the lower limbs assisted by the right upper limb ("hand and hip"), were delivered to the pelvis. Analysis of force and electromyogram recordings from the legs, arm and hand in the hand-and-hip condition showed a close co-ordination of upper and lower limbs in terms of mean latencies and amplitudes. However, trial to trial fluctuations of forces generated by the hand and leg did not correlate, suggesting parallel pathways under central co-ordinative control.

Adult↗
PubMed1999

Impaired anticipatory finger grip-force adjustments in a case of cerebellar degeneration.

We describe adjustments in grip force as a consequence of fluctuations in inertial load force during vertical movements of the upper limb in a patient with cerebellar degeneration. Normally grip force is adapted to load-force fluctuations, in particular to the maximum load force, which occurs early in upward movements and late in downward movements. Increased grip force during movement was observed in the patient, but the timing of maximum grip force was not different between upward and downward movements. This suggests impaired cerebellar prediction of the dynamic consequences of voluntary movement.

Adaptation, Physiological↗
PubMed1994

Ceftriaxone in the treatment of chronic donovanosis in central Australia.

OBJECTIVES: To determine the effectiveness of intramuscular (IM) ceftriaxone sodium in the treatment of chronic donovanosis, and the acceptability to patients and staff of supervised outpatient treatment in rural clinics. METHODS: We collected demographic and sexual health data from participants using a standard questionnaire, and recorded their donovanosis lesions at baseline using genital diagrams. Treatment consisted of a single daily IM injection of 1 g ceftriaxone diluted in 2 ml of 1% lignocaine. Clinic staff followed patients for between three and 12 months, enabling the detection of late recurrences. SETTING: Rural Aboriginal communities in central Australia. PARTICIPANTS: The study describes eight women and four men with chronic donovanosis in detail, and summarises the outcome in 12 additional cases. All cases presented with advanced lesions which had failed to heal on the standard oral antibiotic regimens used in the region. RESULTS: The mean duration of infection was 3.0 years (SD 1.9 years), and between four and ten courses of antibiotics had been prescribed for six of the 12 patients. Patients received between 7-26g of ceftriaxone sodium. Clinical improvement was dramatic in most lesions, and four patients healed completely without recurrence after a total 7-10g of ceftriaxone. Mild recurrences responded to further ceftriaxone or short courses of oral antibiotics. Treatment was well tolerated, and both patient and staff compliance high. CONCLUSION: Donovanosis is an important cause of chronic genital ulceration in central Australia, and is potentially an important risk factor for HIV transmission in Aboriginal communities. The pharmacokinetics and safety profile of ceftriaxone make it a useful and cost-effective agent in the ambulatory management of donovanosis, especially in remote communities. Supervised multidrug regimens of two or more long-acting agents may provide the best answer in donovanosis, administered through the existing health care infrastructure.

Adult↗
PubMed1992

Effects of reducing plantar support on anticipatory postural and intentional activities associated with flexion of the lower limb.

This study examines the anticipatory dynamics of the center of gravity and the EMG activity of ipsi- and contralateral (to the moving leg) soleus (SOL) and tibialis anterior (TA) in flexion of the lower limb executed from different plantar support surfaces. Subjects were standing initially on a plane surface or a surface reduced to two narrow rods under the moving or stance foot. Reduction of the plantar support surface under the moving limb induced a decrease in the anticipatory activity of both TA and in the intentional ipsilateral SOL EMG activity, but the anticipatory activity of SOL did not change. The lateral and vertical accelerations of the center of gravity decreased. When the reduction was under the stance limb, the biomechanical traces did not change markedly and the EMG activity was similar to that of the control situation. The differential effects in EMG activity, depending on whether the reduced plantar surface was under the moving or the stance side, suggest that tonic plantar stimulation induces complex reflex actions on muscles involved in the preparation and execution of voluntary movement. The functional demand could explain the gating from one motor organization to another.

Adult↗