Biomedical subjects
Publications and source records attributed to R Newcomb.
This article provides results of a comparison between the Halstead-Reitan tapper and a 1988 computerized tapping program by Loong. Care should be taken in using the computerized tapper in diagnosis or assessment.
Multiple biologically active peptides arising from a common prohormone are sorted into distinct classes of dense core vesicles within the bag cell neurons of Aplysia californica. In this study, pulse-chase analysis, combined with subcellular fractionation on Percoll gradients, are used to define the location of the prohormone processing events within the secretory pathway. Initial cleavage of the prohormone occurs in a light cellular compartment associated with the Golgi apparatus. The amino-terminal processing intermediate then accumulates in a denser compartment containing small dense cores enclosed in membranous sacs, as well as larger immature vesicles. After 4 h, amino-terminal products are found primarily in a much denser compartment which consists of large and small dense core vesicles. These large and small vesicles can be separated from each other using Percoll gradient centrifugation and are found to be enriched in amino- and carboxy-terminal products, respectively. Lastly, membrane association experiments suggest differential binding to membranes, or integral membrane proteins, as a possible mechanism for sorting of amino- and carboxy-terminal products.
The ELH prohormone is proteolytically processed into at least nine peptides which govern egg-laying behavior in Aplysia. Quantitative immunocytochemistry demonstrates that peptides derived from the prohormone are packaged into distinct vesicle classes. Further experiments suggest the segregation occurs via a rapid initial proteolytic cleavage of the prohormone followed by sorting at the trans Golgi. Egg-laying hormone (ELH) immunoreactivity is localized to the cell body and processes, while bag cell peptide (BCP) immunoreactivity is greater in the cell body. Steady state levels of the amino-terminal set of peptides including the BCPs are 3- to 8-fold lower than the carboxy-terminal cleavage products, such as ELH. Thus, intracellular packaging and routing of the peptides cleaved from a single prohormone regulate their localization and levels in these neurons.
Egg laying in Aplysia is mediated by a battery of neuropeptides released from the bag cell neurons. Predominant intermediates in the proteolytic processing of the Aplysia egg-laying hormone neuropeptide precursor were characterized using biochemical and immunological techniques. Following removal of the signal peptide, a rapid cleavage at the tetrabasic sequence Arg-Arg-Lys-Arg separates the amino and carboxyl regions of the prohormone. Processing of the carboxyl-terminal portion of the precursor then proceeds rapidly via two further cleavages at dibasic residues, resulting in a well defined product mixture within 4 h of chase. By contrast, processing of the amino-terminal side of the molecule proceeds only partially to completion after 20 h of chase and a well defined set of intermediates is not observed. Molecular genetic, physiological, and behavioral studies in conjunction with the biochemical investigations presented here are defining the information flow which governs the egg-laying behavior of Aplysia.
A number of animal behaviors are influenced by the actions of neuropeptides that arise from the processing of complex protein precursors. In this report we investigate the proteolytic processing of neuropeptide precursors expressed in the Aplysia californica bag cells, which govern egg-laying, and neurons R3-14, which mediate aspects of cardiac output. Peptides were purified by fractionation on 2 high-pressure liquid chromatography systems followed by determination of amino acid compositions. Most of these compositions are indicative of processing products derived from the egg-laying hormone (ELH) and R3-14 precursors by cleavage at basic residues. We characterized 9 peptides that arise from the ELH precursor by cleavage of the signal sequence, as well as 7 out of 8 dibasic residues and at least 1 single Arg residue. The peptides range in size from 5 to about 60 amino acids. The R3-14 neuropeptide precursor is cleaved at 2 internal dibasic residues in addition to the signal sequence, resulting in 3 peptides. Shortened forms of several peptides probably result from amino- and carboxy-terminal peptidase action. It is likely that the complex mixtures of neuropeptides arising from these single protein precursors are co-secreted.
In 9 young subjects with mild asthma, we investigated the possibility that chronic daily administration of an inhaled muscarinic antagonist (ipratropium bromide) might increase the response of muscarinic receptors in airway smooth muscle to agonist stimulation caused by receptor upregulation. Subjects inhaled 60 micrograms of ipratropium 4 times daily for 3 wk. Methacholine bronchoprovocation (with or without acute pretreatment with ipratropium) was performed before (control period) and during 3 wk of daily ipratropium therapy (medication period). At the end of the medication period, subjects returned at 12, 24, 48, and 72 h after the last dose of ipratropium (withdrawal period) to determine the provocative concentration of methacholine producing a 20% decrease in FEV1 (PC20). During the medication period, there was no significant diminution of the acute bronchodilator response to ipratropium or of the protective effect of ipratropium against methacholine-induced bronchospasm when compared with the control period. During the withdrawal period, mean in PC20 was significantly less (increased airway responsiveness) at 24 h than during the control period (p less than 0.01) and returned to the control period value within 48 to 72 h. We conclude that daily administration of ipratropium to mildly asthmatic subjects for 3 wk does not produce tolerance to either the bronchodilator effect of ipratropium or to its inhibition of methacholine-induced bronchospasm but does induce transient supersensitivity of airway cholinergic receptors to muscarinic stimulation.
Methods of transferring one bovine embryo to the ipsilateral uterine horn have been compared. In heifers mid-ventral surgical laparotomy under general anaesthesia (n=22) was compared with flank surgery with paravertebral anaesthesia (n=21). Pregnancy rate was higher with a midventral approach (P less than 0.05) (77.3 per cent of 42.9 per cent respectively). In heifers mid-ventral surgery (n=36) was compared with two methods of non-surgical transfer either using the Cassou insemination gun (n=36) or a modification of it (n=39) with embryos collected on days 7, 8 or 10 after oestrus. There were no differences in pregnancy rate between methods or different ages of embryos (methods: 55.6, 55.6, 43.6 per cent respectively; Day: 48.8, 55.8, 48.1 per cent respectively) but the condition score of recipients affected success (P less than 0.05). Of 10 cows which each received an embryo using the modified Cassou gun eight became pregnant.
Fourteen heifers were superovulated five to 10 times using pregnant mare serum gonadotrophin (PMSG) and prostaglandin in a standardised regime which resulted in a mean interval of 42.1 days (SEM +/- 0.97) between treatments. There was a significant (P less than 0.02) fall in the mean ovulation rate at the second treatment when compared with the first but no significant difference between mean ovulation rate at the second and subsequent treatments. No significant difference between treatments in mean egg recovery rate or in the percentage of normal eggs recovered was demonstrated. Four heifers treated 10 times continued to respond with multiple ovulations on most occasions. At the end of the experiment 13 of the 14 heifers were inseminated and 12 conceived, requiring a mean of 1.67 services per pregnancy.
The effect of two dose levels (1000 and 2000 i.u.) of three different commercially available batches of PMSG on the ovarian response (ovulations and follicles greater than 10 mn) of 42 heifers was examined in a randomized incomplete block experiment. Each animal was subjected to two consecutive but different treatments. A significant effect of dose was observed and there were fewer ovulations, but no reduction in the number of follicles, after the second PMSG treatment. There was no evidence that the ovarian response was affected by the PMSG batch used.
Day 7 eggs (oestrus = Day 0) were surgically transferred singly to the uterine horn contralateral to the CL and heifers were either slaughtered on Day 24-26 (Group 1) or palpated per rectum on Day 42 (Group 2) to determine the presence of a developing conceptus and a maintained corpus luteum. There was a fall in pregnancy rate of the control heifers in Groups 1a (6/10) and 2a (2/10) (P = 0.085). Pregnancy rate was unaffected by progesterone treatments from Day 13 to the day before slaughter in Group 1b (8/10), or to Day 23 in Group 2b (2/10). Two of the conceptuses recovered in Group 1b were developing in the presence of regressed corpora lutea. There was, however, an increase in pregnancy rate in Group 2d (6/10) compared with Group 2a (2/10) when heifers were treated with hCG from Days 13 to 35 (P = 0.085). The results indicated that embryonic death is occurring after Day 24 and suggest that a change in the nature of the embryonic signal may be occurring at this time.
A surgical demonstration of the potential use of egg transfer for converting a herd of cattle from one breed to another (Jersey to Friesian) was undertaken on farm. Friesian donors were non-lactating but 50 per cent failed to respond adequately (greater than 3 ovulations) to treatment with PMSG (1500 to 3000 iu). Heifers yielded more ovulations and eggs than cows, but recovery rate was higher from cows (80 per cent cf 60 per cent). Fertilised eggs were recovered from old (13 years) cows, some of which had ceased to breed normally. Pregnancy rate after transfer of these eggs was normal. Purebred Friesian calves out of Jersey recipients were Friesian weight (male, 44.8 kg; female, 37.4 kg) and contrary to experience with crossbreds caused severe dystocia problems. Severity of dystocia was related to birth weight and sex of calf.
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The non-surgical recovery of bovine embryos was examined using a three-lumen PVC catheter passed to the tip of each uterine horn. The recovery of eggs placed in Dulbecco's phosphate buffered saline, with protein was very efficient whether in boiling tubes or funnels. In the absence of protein egg recovery was considerably lower. In a group of 42 heifers, superovulated on two occasions, embryos were recovered non-surgically on day 7 after the first superovulation (Day 0 = oestrus) and surgically after the second. Laparotomy was performed on each occasion and the number of corpora lutea counted. Ovulation rate was lower at the second superovulation and a correspondingly greater recovery of embryos was obtained surgically (P less than 0.02). Non-surgical recovery rates from anaesthetised cows and heifers and from standing sedated heifers were found to be similar. The distance of the catheter tip from the utero-tubal junction (UTJ) significantly affected recovery rate (P less than 0.001). Best results were obtained when the catheter tip was within 5 cm of the UTJ. Most embryos were recovered in the first 100 ml of recovered medium. The method compares favourably with other reported methods.
When transfer of Day 7 embryos was surgically to the horn ipsilateral to the CL and non-surgically to the contralateral horn, fetal survival rate was 85 and 35% respectively. After non-surgical transfer to the ipsilateral and surgical transfer to the contralateral horn, fetal survival rate was 20 and 30% respectively.
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Six heifers with normal oestrous cycles were treated i.m. with 100 i.u. oxytocin on 3 consecutive days, commencing on Days 1-6 after oestrus, and the levels of prostaglandin (PG) F in posterior vena cava plasma were compared with pretreatment values. An increase of PGF in response to oxytocin was significantly influenced by day, with the greatest response occurring on Day 3 after oestrus. In an ovariectomized heifer the levels of PGF in posterior vena cava plasma increased 24 h after priming with oestradiol, but no further increase occurred after oxytocin injection. Peak levels of PGF were higher in the plasma of the posterior vena cava than in the jugular vein. Various storage conditions of the blood before centrifugation and freezing (--20 degrees C) produced significant differences in plasma levels of endogenous PGF, but storage experiments with added labelled PGF-2alpha indicated that the PG was stable in plasma and whole blood.
Although it may ultimately be possible to fertilise ovarian follicular oocytes in vitro, superovulation and insemination are currently the only practical method for obtaining a supply of fertilised ova. It is likely that this will remain true for sometime to come. Pregnant mares serum gonadotrophin (PMSG) is almost universally used for inducing superovulation, and response to it has recently been more reliably achieved by using, in addition, prostaglandin F2alpha to induce oestrus in superovulated donors. The recovery of fertilised ova from donors depends on the expected site of the ova within the genital tract, but both surgical and non-surgical methods involve suspending the ova in a stream of fluid which is collected into glass cups and examined microscopically. A variety of in vivo and in vitro environmental conditions affect the success of transfer of fertilised ova. These include the healthy state of the recovered ovum, the storage medium, the method and length of storage, the age of the ovum and the synchronisation of oestrus of donor and recipient.
A vertical diplopia in a 53-year-old woman was diagnosed as euthyroid Grave's disease, which was arrested by systemic chemotherapy. Subsequent corrective surgery was effective.