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Biomedical subjects

B Engel

Publications and source records attributed to B Engel.

At least 37 records · Page 2Linked to original sources

Gene therapy for pediatric AIDS.

Gene therapy is an experimental treatment modality under investigation for applications to HIV-1 infection. We have developed retroviral vectors carrying anti-HIV-1 genes, demonstrated that these genes cause significant suppression of HIV-1 replication in cultures of primary hematopoietic cells, and performed a clinical trial in pediatric AIDS patients. Four HIV-1-infected children and adolescents underwent bone marrow harvest from which CD34+ cells were isolated and transduced by a retroviral vector carrying an RRE decoy gene. The cells were re-infused into the subjects, without complications, showing that gene transfer in pediatric AIDS patients in safe and feasible. However, gene-containing leukocytes in the peripheral blood were seen only at a low level and only in the first months following cell infusion. To attain some degree of efficacy, it will be necessary to achieve a higher level of gene transfer and to obtain sustained gene expression. We are currently developing new gene transfer methods and vectors designed to improve the results in future trials. If it becomes possible to reach the ideal goal of producing high percentages of T lymphocytes and monocytic cells that are resistant to HIV-1 infection, gene therapy could serve as a complement to antiretroviral drug therapy and help to sustain immunologic function.

Acquired Immunodeficiency Syndrome↗

The effects of selection of pigs on growth rate vs leanness on histochemical characteristics of different muscles.

The microstructure of two type of muscles was studied in a selection experiment conducted with Dutch Large White pigs (boars and gilts) selected for either low backfat thickness (L-line) or fast growth (F-line). Second- and fourth-generation pigs were used to determine effects of selection on fiber type composition, fiber area, and capillary density in the longissimus lumborum (LL) and biceps femoris (BF) muscles. Immediately after slaughter samples were taken from the LL and BF muscles. The latter was divided into an inside (BFi) and outside (BFo) portion, which refer to the red and white portions of the biceps femoris. Serial sections were stained for ATPase (pH 4.60), succinate dehydrogenase (SDH), and alpha-amylase-periodic acid shiff (PAS) to determine fiber type and capillary density. The LL and BFo muscles had predominantly type IIBw fibers, whereas the BFi muscle had a 2 to 4 times higher amount of type I fibers. In most muscles there were more type I and fewer type IIBw fibers in F- than in L-line pigs (P < .05), except in the LL muscle of second-generation pigs and in the red part of the BF muscle of fourth-generation pigs. In both selection lines lower type I and higher type IIBw percentages were found in muscles from gilts than in those from boars (P < .05). Capillary density and fiber area of L- and F-lines showed minor differences, which could be explained by differences in weight and age of the pigs of both lines. The results suggest that selection for low backfat thickness in pig breeding compared with increased growth rate resulted in fewer oxidative and more glycolytic muscle fibers. The magnitude of the effect depended on muscle type and duration of the selection period.

Adenosine Triphosphatases↗

In vitro phagocytosis of boar spermatozoa by neutrophils from peripheral blood of sows.

A considerable number of spermatozoa are used in each sow in routine artificial insemination. However, within a few hours after insemination, many spermatozoa are phagocytosed by polymorphonuclear leucocytes. Some aspects of sperm transport in the female genital tract in the sow have been thoroughly investigated, whereas little is known about the mechanisms involved in the phagocytosis of spermatozoa, or about which spermatozoa (fresh, capacitated or dead) are the most susceptible to ingestion by polymorphonuclear leucocytes. In this study, phagocytosis was investigated by use of an in vitro phagocytosis assay. Polymorphonuclear leucocytes were challenged with either untreated, cold-shocked or frozen-thawed spermatozoa, or with spermatozoa that had been treated to induce capacitation in vitro. The influence of serum on phagocytosis was also investigated. Treatment of the semen to induce capacitation in vitro considerably reduced the phagocytosis of spermatozoa, whereas crude treatments like cold-shock or freezing and thawing reduced phagocytosis only in the first 15-30 min of incubation with polymorphonuclear leucocytes. Viable spermatozoa were phagocytosed mainly through a pathway that was independent of complement or other serum components (for example, antibodies). Complement had little effect on phagocytosis of spermatozoa, but did cause acrosomal exocytosis and cell death.

Acrosome Reaction↗

The importance of surgical clips for adequate tangential beam planning in breast conserving surgery and irradiation.

PURPOSE: To evaluate the role of surgical clips in the planning of tangential beams in patients undergoing breast conserving surgery and adjuvant radiotherapy. METHODS AND MATERIALS: Between September 1996 and April 1998, 25 consecutive female patients with ductal carcinoma in situ, Stage I and II cancer of the breast, underwent lumpectomy with the excision cavity marked by the surgical clips. Subsequently, tangential fields were planned using clinical and radiologic information obtained during simulation without the clips position being visible. RESULTS: When measured from the center of the deepest clip to the posterior field border of the tangential fields in 6/25 patients (24%) the distance was less than 2 cm, with the closest being only 0.5 cm. Respective measurements for the distal end of the clip and the posterior border were: 9/25 patients (36%); less than 2 cm, with the closest distance of 0.3 cm. There was a positive correlation between the distance from the scar to the palpable breast border and the distance between the deepest clip and the posterior border of the tangential beams. CONCLUSION: The lumpectomy scar is often a poor indicator of the excision cavity as demarcated by surgical clips. Without the clips, part of the cavity may be underdosed by the tangential radiation beams. This is more likely for the cavities located close to the lateral or medial border of the breast tissue.

Breast Neoplasms↗

Multimodality diagnostics and megatherapy in poor prognosis Ewing's tumor patients. A single-center report.

BACKGROUND: The prognosis of Ewing's tumor patients has been improved gradually through cooperative therapy studies, so that meanwhile 55 to 65% of the patients survive relapse-free in the long term. Patients with multifocal primary, early or multiply-relapsed Ewing's tumors have a dismal prognosis. Megatherapy with subsequent stem cell transplantation seems to improve outcome in this patient cohort. Feasibility of this intense megatherapy regimen is crucially dependent on collaboration and multidisciplinary coordination of radiologists, radiotherapists, surgeons and oncologists. PATIENTS AND METHODS: Since 1988, 25 patients were treated with megatherapy consisting of melphalan, etoposide and total-body irradiation followed by stem cell transplantation. All patients received 6 courses of an induction therapy. Before the fourth therapy block, tumors that were bulky at initial diagnosis (> 200 ml) were excised, as well as lung metastases which could still be detected after the third chemotherapy block. During the fifth and sixth chemotherapy block, patients received local irradiation on all infiltrated sites. Persisting immunosuppression after high-dose treatment may facilitate the incidence of relapse. To improve proliferation and cytotoxic activity of early regenerating NK-cells, adoptive immunotherapy with systemic IL-2 therapy after megatherapy is part of the treatment protocol. RESULTS: Of 25 patients treated with megatherapy, 10 patients are still alive with a follow-up time of 6 months to 9 years after megatherapy. The time up to engraftment was decreased from 15 +/- 6 days down to 9 +/- 2 days through the use of G-CSF and CD34+ selected stem cells. At the same time, erythrocyte and platelet replacement was shortened. Frequently occurring complications were mucositis and infections. One patient died after developing septicemia and multi-organ failure, another patient developed a myelodysplastic syndrome 4.5 years after megatherapy. However, relapse is still the major cause of death. The influence of IL-2 on event-free survival cannot valued because 21 of 25 patients were treated with adoptive immunotherapy. CONCLUSION: The complex diagnostic and therapeutic strategy renders and EFS of 34% for a patient group with otherwise dismal prognosis. To clarify the efficiency of megatherapy in patients with advanced Ewing's tumors, a standardized treatment strategy is necessary to accumulate a sufficient number of patients for large cooperative studies in this subject.

Adolescent↗

The effect of feed withdrawal on broiler blood glucose and nonesterified fatty acid levels, postmortem liver pH values, and carcass yield.

Feed withdrawal should start 5 h before broilers are captured and transported to the processing plant. This practice results in a total feed withdrawal period of about 8 h before slaughter. To check observance of this period at the plant reliably, blood samples were collected at slaughter from broilers. These samples were analyzed for glucose and nonesterified fatty acid (NEFA). Glucose levels decreased, whereas the amount of NEFA increased as a consequence of deprivation. It was found that the decrease in glucose was smaller than could be expected from data in the literature, whereas the level tended to rise again after longer deprivation periods. The increase of the NEFA was lower in males than in females, and the glucose: NEFA ratio also showed a difference between the sexes. Therefore, it was concluded that these blood metabolites were not suitable for use as an indicator of the duration of feed withdrawal. The estimation of liver pH seemed to be a more reliable indicator of the length of feed withdrawal. Longer periods of feed withdrawal resulted in higher ultimate liver pH values, which were 6.10 to 6.20 in full-fed broilers, increasing to 6.60 in livers from broilers deprived during longer withdrawal periods. Further research, however, is needed because broilers in the present study were exposed to minimal amounts of stress. Other carcass characteristics such as slaughter, evisceration, and oven-ready yields matched results previously described in the literature.

Abattoirs↗

Influence of capacitation and fluids from the male and female genital tract on the zona binding ability of bull spermatozoa.

Before fertilization, inseminated spermatozoa acquire the ability to fertilize an egg, a phenomenon called capacitation. Bovine sperm capacitation is influenced by factors originating from both the male and female genital tract, and results in intracellular and membrane changes of the spermatozoa that facilitate the induction of the acrosome reaction. However, the effects of reproductive tract secretions and capacitation on the binding of spermatozoa to the zona pellucida have not been investigated. In this study, a sperm-egg binding assay was used to determine whether the ability of bull spermatozoa to bind to the zona pellucida was altered during in vitro capacitation by heparin or oviductal fluid, or by treatment of spermatozoa from the cauda epididymidis with accessory sex gland fluid. In addition, biotinylated solubilized zona pellucida proteins were used to visualize zona binding on spermatozoa. The ability of bull spermatozoa to bind to the zona pellucida was increased after both heparin and oviductal fluid induced in vitro capacitation. Exposure of spermatozoa from the cauda epididymidis to accessory sex gland fluid resulted in a direct increase in zona binding ability, followed by a further increase during capacitation in vitro. Binding of solubilized zona proteins was restricted to the acrosomal cap of bull spermatozoa. It is suggested that the observed increased ability of bull spermatozoa to bind to the zona pellucida enables optimal sperm-egg attachment, which also relates to the induction of the acrosome reaction by the zona pellucida. Thus, increased zona binding ability is likely to be an essential part of the process of capacitation.

Animals↗

Effects of trehalose and sucrose, osmolality of the freezing medium, and cooling rate on viability and intactness of bull sperm after freezing and thawing.

The influence of the presence of trehalose or sucrose, the osmolality of the freezing medium, and the cooling rate on success of cryopreservation of bull sperm were investigated. Bull semen was frozen at four different cooling rates varying from 40 to 300 degrees C/min in standard Tris-egg yolk medium and in hypertonic or isotonic media containing 0.2 M trehalose or sucrose. Sperm motility, H33258 exclusion, and acrosome morphology were evaluated. In the hypertonic media the motility was almost completely inhibited, but could partly be restored by returning to isotonic conditions. Medium composition and cooling rate had significant effects, with a strong interaction of medium x cooling rate. The optimal cooling rate was found to be between 76 and 140 degrees C/min. At the highest cooling rate, both the presence of the sugars and the high osmolality of the hypertonic media conferred significant protection against fast-cooling damage, sucrose being significantly better than trehalose. At all other cooling rates the medium differences were less pronounced, but the isotonic sugar media were generally superior to standard medium as to preservation of acrosome intactness and H33258 exclusion. This indicates that the isotonic sugar media could enable some improvement of cryopreservation of bull semen over the currently used Tris-egg yolk medium. Further improvement of sperm survival at the AI station should be possible by implementation of controlled rate freezing, using a cooling rate in the optimal range between 76 and 140 degrees C/min.

Animals↗

The circadian rhythm of salivary cortisol in growing pigs: effects of age, gender, and stress.

This experiment was designed to examine circadian rhythmicity of cortisol in saliva of growing pigs, in relation to age, gender, and (time of) stressor application. Additionally, the acute cortisol response to a stressor was studied. Five groups, each consisting of 3 barrows and 3 gilts, were involved in the experiment. In a Control Group, saliva samples were taken at 1-h intervals at 12, 16, 20, and 24 weeks of age. Within 1 week, rhythmicity of cortisol was assessed during two 24-h spans (Monday and Friday). Rhythm characteristics were evaluated by cosinor analysis, describing the rhythm by several parameters. In 2 groups at 12 weeks and 2 other groups at 20 weeks of age, a stressor was applied (4 h of isolation) on Thursday morning or evening. Again, rhythmicity was assessed on Monday and Friday by sampling at 2-h intervals. Acute cortisol effects were studied by sampling at several time-points during isolation. Between 12 and 24 weeks of age, basal cortisol concentrations decreased and a rather stable and adult circadian rhythm was reached at 20 weeks of age. Average basal cortisol concentrations were higher in barrows than in gilts. Furthermore, after isolation, the amplitude of the rhythm was increased in barrows but was unchanged in gilts. The rhythm was more unstable and the maximum value tended to shift only after evening isolation. Stressor timing, but also age, was found to affect average cortisol concentrations. Moreover, stressor timing was important for the acute cortisol response: the increase was higher in the morning. The results of this study emphasize the importance of considering the circadian rhythmicity of cortisol, in relation to age, gender, and (time of) stressor application, when studying the cortisol response of animals to stressors.

Age Factors↗

Predicting optimal time of insemination in cows that show visual signs of estrus by estimating onset of estrus with pedometers.

An experiment was designed to estimate the optimal interval from the beginning of estrus to artificial insemination (AI). The data were analyzed by means of a mathematical model. The analysis was based on pedometer readings and results of rectal palpation at 42 to 49 d post-AI of 171 breedings in 121 cows. The chance of conception was highest between 6 and 17 h after increased pedometer activity; the estimated optimum was at 11.8 h. In this data file, the effects of disease, inseminator, time of AI (a.m. or p.m.), and bull did not contribute to the improvement of the model. The effects of disease were not significant because of the low incidence of any specific disease. Activity measurements can be used as a tool for AI strategy to improve conception in groups of healthy cows and heifers already showing visual signs of estrus.

Animals↗

Maximizing selection efficiency for categorical traits.

Genetic improvement of categorically recorded traits is hampered because information content of categorical records is low and ordinary linear breeding value estimation methods do not apply theoretically. The ordinary animal or linear mixed model (LMM), which ignored the categorical nature of the trait, is compared to a generalized linear mixed model (GLMMp) that assumes a linear mixed model for an underlying continuous variable. The GLMMp takes full account of the categorical nature of the trait and is a straightforward extension of LMM. In a closed nucleus breeding scheme (e.g., cattle, pigs, or poultry), rates of genetic gain increased by 1 to 2%, when GLMMp was used instead of LMM. Rates of genetic gain increased by 7 to 20%, when the best sires were used on the best herds (i.e., when there was some confounding between sire and herd effects). When considering a binary trait (e.g., disease incidence) initial incidences of 25% could be reduced to 2.8% within 10 generations of selection. Rates of gain can be increased by up to 84% by gathering more information on high-incidence categories (i.e., by dividing these categories into subcategories). Subdividing low-incidence categories (e.g., splitting diseased animals into moderately and severely diseased) hardly increased rates of gain. Direct recording of the underlying variable, which requires uncovering of the physiological background of the categorical trait, yielded 109 to 278% more genetic gain than selection for a binary trait.

Animals↗

[Recent progress in therapy of the lacrimal ducts 1991].

We went through papers on lacrimal duct disorders published in 1991 seeking new information which could be directly applied. Practical clinical information was given priority over academic results. 1. New strategies were adopted in particular in the surgical treatment of lacrimal duct obstructions. The most important progress in this context was in the use of various types of laser in combination with improved video endoscopes, e.g. in laser dacryocystorhinostomy or laser conjunctivodacryocystorhinostomy. 2. A special instrument, the "Reiss punctal punch", was developed for treatment of punctal stenosis. It enhances and facilitates punctoplasty. 3. With regard to the lacrimal gland and tear secretion, it was shown that epidermal growth factor (EGF) is a natural component of human tears and is formed in the lacrimal gland. It is not yet known whether corneal wound healing can be influenced by administering drugs which alter EGF production.

Adolescent↗

Analysis of embryonic development with a model for under- or overdispersion relative to binomial variation.

Observations from a study of the development of ovulations into embryos for Texel sheep are analysed with a model for count data that are under- or overdispersed relative to binomial variation. The analysis is based on maximum quasi-likelihood (McCullagh and Nelder, 1989, Generalized Linear Models, 2nd edition, London: Chapman and Hall), following an approach suggested by Williams (1982, Applied Statistics 31, 144-148). The dispersion parameter is developed as a combination of a variance component representing shared maternal effects and a correlation, typically negative, between ovulations within ewes. The number of ovulations (the binomial denominator) is included as an explanatory variable.

Androstenedione↗

Development of a high capacity radioimmunoassay procedure: ovine follicle stimulating hormone determination in blood plasma as a model.

A sensitive, specific and accurate homologous radioimmunoassay (RIA) for ovine follicle stimulating hormone (oFSH) has been developed, using a [125I]oFSH tracer and a polyclonal rabbit anti-OFSH-serum at a final dilution of 1:224,000. The separation of free and antibody-bound tracer is based on the double antibody solid phase system. The assay was found to be specific for oFSH; cross-reactivity with oLH, oPrl and oGH was lower than 0.2%. The detection limit was 7 pg per tube. Inter- and intra- assay coefficients of variation (CV) were 3.7-8.5% and 6.6%, respectively. The use of a few selfmade appliances in combination with a well-considered time-schedule enabled the processing of three thousand blood plasma samples in triplicate within two weeks. The particular advantage of the method described here is the fast, easy and safe separation of free and antibody-bound tracer with minimal handling of radioactive tubes.

Animals↗