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Biomedical subjects

L Manley

Publications and source records attributed to L Manley.

At least 19 recordsLinked to original sources

Re-examining steroid hormone metabolites as ovulation markers using monoclonal antibodies.

Plasma progesterone and plasma estradiol levels are commonly used to monitor ovulation in women although for the adequate documentation of ovulation the expense and discomfort of multiple venipuncture sampling may be required. Accurate and definitive information on ovulation in women can be obtained by the simple measurement of metabolites of progesterone and estradiol in early morning urine samples. These analyses have been made possible by the use of our own highly specific monoclonal antibodies to both pregnanediol-3-glucuronide (P-3-G) and estrone-3-glucuronide (E-3-G) and the development of simple, direct, automated enzyme-linked immunosorbent assays. Sequential sampling and the generation of ovulation profiles allows detection of ovulation and identification of the infertile/fertile phases of the cycle for either planned pregnancies or natural family planning. Aberrations in ovulation are easily detected as is documentation of the transition to menopause. The use of sequential, spaced, early morning urine samples for P-3-G and E-3-G allows accurate assessment of ovulatory function rather than relying on the usual single plasma sampling. The data presented also show that the direct determination of plasma pregnanediol-3-glucuronide in plasma is as informative as plasma progesterone measurement.

Adult

Production of a monoclonal antibody to cortisol: application to a direct enzyme-linked immunosorbent assay of plasma.

Cortisol mouse monoclonal antibodies were produced and characterized. Of the four clones studied, supernatant from one clone (A2), compared with other cortisol monoclonal antibodies, showed minimal cross-reactivity to other C21 steroids and was suitable for the direct determination of cortisol in plasma by enzyme-linked immunosorbent assay using a standard 96-well microtiter plate. The enzyme-linked immunosorbent assay uses the immobilized antigen approach, in which cortisol in plasma samples or standards competes with immobilized steroid for antibody-binding sites. After washing, the cortisol antibody bound to the wells of the microtiter plate is detected with antimouse immunoglobulin conjugated to horseradish peroxidase. Following further washing, o-phenylenediamine substrate is added. The enzyme-linked immunosorbent assay is robust and semiautomated. The mean +/- SD recovery from plasma was 97% +/- 6%. Precision studies on three different plasma pools showed mean coefficients of variation of 7.6% and 8.6% for within- and between-assay variation, respectively. The satisfactory performance criteria allow its use in the routine laboratory.

Animals

Trauma in pregnancy: uterine rupture.

Trauma during pregnancy is a unique situation. Understanding injury patterns, anatomic and physiologic changes, and the initial approach to resuscitation is essential for all emergency nurses. Maternal resuscitation is the only means of fetal resuscitation. Meticulous attention must be given to the ABC's, with some minor modifications. Cervical spine immobilization is done in conjunction with positioning on the left side. Oxygen is used liberally, but may not benefit the fetus if hypovolemia exists. IV access and aggressive fluid resuscitation should proceed quickly. Diagnostic testing, including radiologic evaluation, is performed as necessary--the mother's life must not be jeopardized on the basis of fetal risk. Continuous fetal monitoring should be instituted, even with seemingly minor injuries. In the rare event of maternal arrest, a postmortem cesarean section may be lifesaving for the infant. Policies should be formulated jointly by ED, obstetric, and neonatal staffs in advance to speed this difficult decision-making process. The keys to survival, for both mother and infant, are an organized approach to resuscitation and teamwork among all professionals.

Adolescent

Use of a monoclonal antibody to estrone-3-glucuronide in an enzyme-linked immunosorbent assay (ELISA).

A direct urinary ELISA for estrone-3-glucuronide has been produced following cloning and characterisation of a monoclonal antibody to the above estrogen metabolite. The ELISA follows our established pattern of absorbing a thyroglobulin conjugate, to which estrone-3-glucuronide has been coupled, to the wells of a microtitre plate using guanidine hydrochloride. A competition reaction between either standards/samples and the adsorbed hormone compete for antibody combining sites. The assay is completed by addition of an anti-mouse Ig-peroxidase complex and read at 492 nm following additions of O-phenylenediamine substrate in under 4 h. The correlation between urinary "total estradiol" and "total estrone and estradiol" is very good and, in conjunction with our ELISA for pregnanediol glucuronide, has allowed for the improved clinical management of infertile and subfertile women.

Animals

Apnoeic heart rate responses in humans. A review.

Research examining the heart rate response of humans to apnoea and apnoeic immersion in water has tended to produce equivocal results. Controversy exists in relation to the existence and onset, course and ultimate extent of apnoeic bradycardia, either at rest or during exercise. The reduction in heart rate that has been observed in response to apnoea and apnoeic immersion is considered by some to be a component of an oxygen conserving mechanism termed the mammalian dive reflex. Doubt has been expressed, however, as to whether humans do possess the complex cardiovascular response evident in diving mammals. A bradycardial response to apnoea at rest is reasonably well established. Reports conflict with regard to apnoeic responses to exercise in both terrestrial and aquatic environments, and for the temporal onset of apnoeic bradycardia in general, and the time course required for the maximal development of the response. It is suggested that the existing discrepancies with regard to heart rate responses to apnoea are due in part to the large variety of research protocols employed, with, in many cases, a lack of control of those factors said to modify apnoeic heart rates, and in part to a lack of uniformity in the interpretation of the obtained results. Conflicting evidence exists for almost all of the factors said to modify the apnoeic heart rate response in humans. Factors implicated in the phenomenon of apnoeic bradycardia include the influence of temperature and the physical condition of the individual, varying lung volumes, the depth of immersion in water, the body position during the apnoeic episode, and the psychological state of the individual. The influence of the sex and age of the subjects is equally contentious. Finally, the role of breath-holding capacity as it influences the extent of bradycardia remains to be examined further. Additional research requires the adoption of an integrative, holistic approach if a comprehensive understanding of apnoeic heart rate responses is to be achieved.

Adolescent