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Biomedical subjects

T J Lam

Publications and source records attributed to T J Lam.

At least 91 records · Page 5Linked to original sources

Evaluation of a prototype on-line electrical conductivity system for detection of subclinical mastitis.

A prototype on-line system for measurement of electrical conductivity of quarter milk was evaluated for accuracy in detection of subclinical mastitis compared with that of bacteriological culture and SCC of sampled quarters. Because of the low quarter prevalence of mastitis, quarters were sampled conditionally from the signals of the on-line system. All signaled quarters and a random selection of the nonsignaled quarters were sampled. To calculate sensitivity and specificity, the total number of non-signaled quarters was extrapolated. The system identified correctly 18 out of 23 subclinical quarters and 521 out of 555 healthy quarters. Quarter prevalence was about 1%. Predictive value of a positive test (35%) and the predictive value of a negative test (99%) were calculated, as well as sensitivity (25%) and specificity (99%), after extrapolation of the total number of nonsignaled quarters. Because of repeated measurements, sensitivity may be underestimated. When signaled quarters were defined by repeated signals within 14 d, predictive value positive increased to 48%. The prototype on-line system did not detect subclinical mastitis very accurately because of suboptimal test characteristics.

Animals↗

Two major groups of vitellogenin cDNA clones from Oreochromis aureus (Steindachner).

cDNA libraries were constructed using livers of male and female Oreochromis aureus that were stimulated with estradiol. Restriction map analyses of vitellogenin (Vg) cDNA clones indicated 4 subgroups with related restriction patterns, represented by pOAVg2 (from female) and pOAVg50, 71 & 87 (from male) fish. With the exception of pOAVg2, all other clones obtained from female fish resembled pOAVg50. There are no internal repeats within these cDNA clones. Southern blot cross-hybridisations distinguished pOAVg87 (group A) from pOAVg2, 50 and 71 (group B). The existence of these 2 major groups of Vg cDNAs was further confirmed by dot blot hybridisations and hybrid melting assays under varying stringencies. The difference of melting temperature, Tm between the 2 groups suggests a 7% intraspecific divergence in the Vg gene family.

Animals↗

[Analgesics; the use of aspirin in dogs; effects of tablet type and food intake on plasma salicylate level].

Administration of acetylsalicylic acid (aspirin) in the dog may cause gastric mucosal damage. Enteric-coated tablets protect the canine stomach during oral aspirin medication. A therapeutic plasma salicylate concentration can be attained using enteric-coated aspirin tablets at a dosage of 25 mg/kg TID. In a series 4 of experiments using adult beagle and large mixed breed dogs and two types enteric-coated tablets, the influence of food intake on the plasma salicylate concentration was studied. Tablets were administered with 8h intervals and food intake was either once daily or three time daily with 8h intervals. Plasma salicylate concentrations were also studied during fasting. It is concluded that, when using enteric-coated tablets, the plasma salicylate concentration in the dog after oral medication is strongly influenced by the aspirin dosage, the tablet type and the feeding pattern. Large enteric-coated tablets may accumulate in the stomach over several days and are not suitable for use in the dog. The gastric accumulation is caused by the enteric-coating of the large tablets and not by the aspirin medication.

Animals↗

Within-herd and between-herd variation in diagnosis of clinical mastitis in cattle.

In a set of data on farmer-observed clinical mastitis in cattle, all milk samples were scored with regard to milk color and milk texture (presence of flakes and viscosity). During a 1-year period, 1,106 milk samples were collected from 125 farms with a bulk milk somatic cell count < 150,000 cells/ml. The overall variation, the between-herd variation, and the within-herd variation of the scores were evaluated and compared with each other. Differences between farmers were found for the scores on the visual characteristics per sample. However, these differences were not related to the reported number of cases of clinical mastitis per year. It was concluded that the diagnostic capability of farmers whose herds had low somatic cell counts did not have a negative influence on the validity of farmer-observed clinical mastitis incidence. However, care should be taken in herds with a low frequency of cases.

Analysis of Variance↗

[Dermatitis digitalis? Considerations in the evaluation of an intensive therapy].

A piece of clinical research was performed to illustrate the recovery process of Digital Dermatitis on a chronically contaminated cattle farm. The combined effect of intensive individual treatment and an antibiotic foot-bath was examined. In addition to the classical condition recognized as Mortellaro's disease we found an aspecific form of Digital Dermatitis which failed to respond adequately to the currently recommended therapy.

Animals↗

Antibodies against nematodes in serum, milk and bulk milk samples as possible estimators of infection in dairy cows.

Antibodies against Ostertagia spp., Cooperia spp. and Dictyocaulus viviparus were assessed in serum and individual milk samples of cows and in bulk milk samples from the refrigerating tank. Titres in milk samples were related to serum titers, but also influenced by milk yield, lactation stage and age of the cow. Correlation coefficients between serum and individual milk samples were highly significant, and varied from 0.42 to 0.56. Correlations between herd means of serum titres and herd means of individual milk titres varied from 0.41 to 0.61, and those between herd means of serum titres and means of bulk milk samples from 0.43 to 0.67. Finally, the correlations between herd means of individual milk samples and means of bulk milk samples varied from 0.52 to 0.82. This variation was dependent upon worm species and whether or not correction for milk yield was applied. The discrimination of herds was evaluated by the between/within herd ratio of variances. F-values for all types of samples were highly significant, and at least as high for milk samples as for serum samples. If, as has been shown for serum samples, a positive relationship exists between the milk yield response to anthelmintic treatment and the mean herd titre of milk samples or bulk milk samples, assessment of these titres could offer a rational basis for treatment of dairy cows.

Age Factors↗

Steroid metabolism by ovarian follicles and extrafollicular tissue of the guppy (Poecilia reticulata) during oocyte growth and gestation.

In the viviparous guppy, fertilization and gestation are intrafollicular. Fully developed embryos are ovulated at the end of gestation just prior to parturition. The metabolism in vitro of various radiolabeled steroid precursors by isolated ovarian follicles at various stages of the reproductive cycle and extrafollicular (EF) tissue of the guppy was investigated. While estradiol-17 beta was one of the end products of metabolism in vitellogenic follicles, 17 alpha, 20 beta-P and several 5-reduced metabolites were synthesized by postvitellogenic follicles. The yield of 17 alpha, 20 beta-P, however, was much lower than some 5 beta-reduced metabolites synthesized by postvitellogenic follicles. Gestation stage follicles rapidly converted the precursors into 5-reduced and polar 7-hydroxylated steroids, and their glucuronides. Although postpartum follicles showed very poor potential for steroid metabolism, they synthesized estradiol-17 beta from testosterone. These results demonstrate distinct changes occurring in the steroidogenic potential of the follicles during the reproductive cycle. Unlike in other viviparous vertebrates, no particular steroid seems to be involved in maintaining gestation in the guppy; all the steroid precursors are converted into highly polar metabolites and their conjugates during gestation, thereby facilitating their excretion. The EF ovarian tissue also synthesized 7-hydroxylated steroids and their glucuronides, providing evidence for the first time that the teleost ovarian EF tissue plays a role in steroidogenesis. The possible physiological significance of the synthesis of the novel polar steroids by the follicles and the EF tissue is discussed.

17-alpha-Hydroxyprogesterone↗

Steroid production by ovarian follicles of the viviparous guppy (Poecilia reticulata) and its regulation by precursor substrates, dibutyryl cAMP and forskolin.

Production in vitro of estradiol-17 beta, testosterone, 17 alpha-20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-P), 17 alpha-hydroxyprogesterone, and progesterone by follicles of the guppy at various stages of oocyte growth and gestation was investigated. Basal production of estradiol-17 beta was highest in 0.8- and 1.2-mm follicles, whereas that of testosterone and 17 alpha,20 beta-P was highest in 1.6-mm (postvitellogenic) follicles. Levels of these steroids declined after fertilization and were undetectable in late gestation and postpartum follicles. 17 alpha-Hydroxyprogesterone and progesterone levels were low at all stages. Thus, none of these steroids appears to be involved in maintaining gestation. Regulation of estradiol-17 beta and 17 alpha,20 beta-P secretion by vitellogenic (1.0 mm) and postvitellogenic follicles by precursor substrates, dbcAMP (0.1 to 10 mM) and forskolin (1 to 100 microM), was also investigated. Vitellogenic follicles synthesized increased quantities of estradiol-17 beta in the presence of exogenous testosterone, whereas estradiol-17 beta production by postvitellogenic follicles was not altered by testosterone. These results suggest decreased aromatase activity in the postvitellogenic follicles. Dibutyryl cAMP and/or forskolin stimulated testosterone and estradiol-17 beta production by vitellogenic follicles but did not stimulate conversion of testosterone to estradiol-17 beta, suggesting that the adenylate cyclase system stimulates estradiol-17 beta production by stimulating testosterone production but does not mediate conversion of testosterone to estradiol-17 beta. Postvitellogenic follicles synthesized increased quantities of 17 alpha,20 beta-P in response to 17 alpha-hydroxyprogesterone in a dose-dependent manner. Although 1 microM of forskolin stimulated 17 alpha,20 beta-P production by postvitellogenic follicles in the absence of exogenous 17 alpha-hydroxyprogesterone, 100 microM of forskolin inhibited 17 alpha,20 beta-P production. Dibutyryl cAMP, however, did not affect 17 alpha,20 beta-P production. In the presence of 50 ng of 17 alpha-hydroxyprogesterone, dbcAMP (10 mM) and forskolin (1 to 100 microM) suppressed 17 alpha,20 beta-P production. It is suggested that cAMP mediates 17 alpha,20 beta-P production up to a certain threshold level, beyond which it inhibits 17 alpha,20 beta-P production.

17-alpha-Hydroxyprogesterone↗

Effects of neurohypophyseal and adenohypophyseal hormones, steroids, eicosanoids, and extrafollicular tissue on ovulation in vitro of guppy (Poecilia reticulata) embryos.

In the viviparous guppy, oocyte maturation is followed by intrafollicular fertilization and gestation. The fully developed embryos are ovulated at term just prior to parturition. Various agents were tested in vitro for their effects on ovulation of embryos in isolated follicles of the guppy. Arachidonic acid (10 and 100 microM), PGE2, PGF2 alpha, and 6-keto-PGF1 alpha (0.1 microgram/ml) induced ovulation, while PGE1, 15-keto-PGF2 alpha, leukotriene B4, 5-, 12-, and 15-HETEs (0.01 to 0.1 microgram/ml), cortisol, 11-deoxycortisol (25 and 250 ng/ml), estradiol-17 beta, testosterone, 17 alpha,20 beta-P, progesterone (5 and 50 ng/ml), isotocin, vasotocin (0.02 to 2 microgram/ml), and guppy pituitary extract (one and two glands per fish) did not. Extrafollicular (EF) ovarian tissue cocultured with isolated follicles induced ovulation, and the medium levels of PGE and PGF in such incubations were higher than those in the control. Indomethacin, the cyclooxygenase inhibitor, did not inhibit ovulation induced by arachidonic acid and EF tissue, although it inhibited PGE and PGF production. NDGA, the lipoxygenase inhibitor, did not inhibit ovulation induced by arachidonic acid or EF tissue. A combination of eicosanoids synthesized by follicles and EF tissue may be involved in the induction of ovulation. Dibutyryl cAMP inhibited ovulation induced by PGE2, PGF2 alpha, 6-keto-PGF1 alpha, and EF tissue suggesting that a low level of cAMP may be associated with ovulation in the guppy.

Animals↗

Prostaglandins and teleost neurohypophyseal hormones induce premature parturition in the guppy, Poecilia reticulata.

Administration of PGE2, PGF2 alpha, isotocin, and vasotocin to pregnant guppies during late gestation induced premature parturition. Indomethacin (10 micrograms/g body wt), the cyclooxygenase inhibitor, did not inhibit parturition at term, suggesting that some lipoxygenase metabolites may be involved in the induction of parturition. Pituitary extract, as well as LH-RH, did not induce premature parturition. It may be that in the guppy, in which ovulation occurs at the end of gestation just prior to parturition, gonadotropin is not involved in the induction of ovulation and parturition.

Animals↗

Steroid metabolism in teleost gonads: purification and identification of metabolites by high-performance liquid chromatography.

A simple, efficient, and comprehensive technique for the purification, identification, and quantitation of the common steroid metabolites synthesized by the gonads of teleosts involving five systems of high-performance liquid chromatography (HPLC) was developed. Steroid standards were identified in HPLC by UV absorption at 254 nm or 280 nm, by differential refractive index, or by using radioactive standards. Metabolites that do not absorb UV light and are not resolved in the isocratic HPLC systems were identified in thin-layer chromatography following purification by HPLC. By using this technique, most of the steroid metabolites, including some polar metabolites, synthesized by the gonadal tissues of the teleosts can be purified within three steps of chromatography. The HPLC systems reported here are also useful in identifying the chromium trioxide oxidized products of metabolites, such as triols and tetrols, which considerably narrows down the number of probable metabolites.

Animals↗

Progestins and cortisol delay while estradiol-17 beta induces early parturition in the guppy, Poecilia reticulata.

Fertilization and gestation are intrafollicular in the guppy (Poecilia reticulata), and ovulation occurs at the end of gestation prior to parturition. In this study, the effects in vivo of the ovarian steroids, progesterone, 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-P), cortisol and estradiol-17 beta, the antiprogestin RU 486, and aromatase inhibitor, 4-hydroxyandrost-4-ene-3,17-dione (4-HAD), on gestation and parturition were studied in the guppy. Progesterone (0.05 and 0.10 micrograms/ml of water), 17 alpha,20 beta-P (0.01 micrograms/ml and greater), cortisol (0.10 micrograms/ml) and 4-HAD (0.10 micrograms/ml) all prolonged gestation presumably by inhibiting ovulation. 17 alpha,20 beta-P was most effective in inhibiting ovulation and parturition for up to 36 days postpartum. This inhibition was reversed when fish were transferred to steroid-free water. Besides extending gestation, 17 alpha,20 beta-P and 4-HAD also inhibited development of vitellogenic oocytes. Estradiol-17 beta (0.05 and 0.10 micrograms/ml) and RU 486 (10 micrograms/g body weight) both induced premature parturition presumably by accelerating onset of ovulation. These results, together with our previous observations on the steroid profile in the guppy, strongly suggest roles for estradiol-17 beta and cortisol in regulating ovulation and parturition.

Animals↗

Effect of thyroid hormones on hatching in the tilapia, Oreochromis mossambicus.

Incubation of fertilized eggs of Oreochromis mossambicus in media containing varying doses of T3, T4, and phenylthiocarbamide (PTC) indicated that the thyroid hormones delayed hatching, while the antithyroid drug accelerated it. Denuded eggs were also incubated in the above media and the media were assayed at various time intervals for hatching enzyme. The thyroid hormones delayed hatching enzyme release, while PTC enhanced it. The possible role of thyroid hormones in the control of the hatching process in fish eggs is discussed.

Animals↗

Estradiol-induced vitellogenin gene expression in a teleost fish, Oreochromis aureus.

Vitellogenesis presents a versatile model for the study of hormone-induced gene expression. We report here the effects of estradiol-17 beta-propionate on vitellogenin gene expression in male Oreochromis aureus, a teleost fish. Vitellogenin mRNA of 6500 nucleotides has been elucidated from the livers of female and estradiol-treated male O. aureus. By hybridization with a specific O. aureus cDNA probe, the vitellogenin mRNA transcript was detected as early as 1 hr following primary and secondary estradiol-stimulations, although for the latter, the rate of accumulation of vitellogenin-specific mRNA was 20-fold higher. The vitellogenin mRNA peaked at 72 and 48 hr, respectively, for primary and secondary stimulations. At the translational level, the increase in plasma vitellogenin was further enhanced during the secondary stimulation. There was a distinct shift in the peak of plasma vitellogenin from Day 14 in the primary induction to Day 3 in the secondary stimulation. The plasma vitellogenin presented in two forms, 300 and 500 kDa, both of which were immunologically confirmed by Western blot analysis to be vitellogenin proteins.

Animals↗

Steroid hormone profile during gestation and parturition of the guppy (Poecilia reticulata).

The guppy is an ovoviviparous teleost with fertilization and gestation occurring intrafollicularly, and ovulation and hatching at around parturition. The levels of estradiol-17 beta (E), testosterone (T), cortisol (F), 17 alpha-hydroxyprogesterone (17-OHP), 17 alpha-20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P), and progesterone (P) in the guppy at different stages of gestation were measured in 60 microliters of serum with a highly specific, sensitive, precise, and accurate procedure involving high-performance liquid chromatography and radioimmunoassay. Levels of E and T increased during vitellogenic growth of oocytes and declined sharply after fertilization, increasing again gradually during late gestation to reach prefertilization levels at periparturition. F levels decreased significantly during fertilization followed by an increase during gestation and a sharp decline again at periparturition. The level of 17-OHP was significantly lower at periparturition than at other stages, while that of 17,20 beta-P was significantly higher in periparturition than in late gestation. There was no significant change in the levels of P at all stages of gestation, suggesting that, unlike mammals, P may not be involved in maintaining gestation in this fish. Changes in the levels of E and T seem to be related to oocyte growth rather than gestation. The role of F in gestation is uncertain since the effect of stress could not be discounted or defined in this study. Thus, on the present results, the role of any particular steroid in maintaining gestation of the guppy is not obvious. The elevated levels of E at term may facilitate parturition in addition to its role in vitellogenesis during oocyte growth.

Animals↗

Gastric retention of enteric-coated aspirin tablets in beagle dogs.

Administration of acetylsalicylic acid (ASA) in the dog may cause gastric mucosal damage. Enteric-coated tablets protect the canine stomach during oral ASA medication. A therapeutic plasma salicylate concentration can be attained using enteric-coated ASA tablets at a dose rate of 25 mg/kg body wt, administered every 8 h. Six beagle dogs were given enteric-coated ASA tablets (500 mg) orally, in a 5-day cross-over experiment on two different feeding regimens: i.e. feeding once daily (Group I) or 8 hourly (Group II). Results demonstrate that feeding regimen strongly influences the plasma salicylate concentration pattern. Subtherapeutic mean plasma salicylate concentrations were found in both groups. In Group II the standard deviation (SD) of the mean plasma salicylate concentration was larger than that of Group I. The minimal plasma salicylate concentration never reached detectable levels in Group II. In both groups large numbers of tablets were vomited. Gastric evacuation of the ASA tablets is comparable to indigestible solid particles; their removal was dependent on the interdigestive gastric motility. It is concluded that large enteric-coated ASA tablets are not suitable for therapeutic use in small dogs.

Administration, Oral↗

Cloning, characterisation and expression of vitellogenin gene of Oreochromis aureus (Teleostei, Cichlidae).

cDNA synthesised from liver Poly A+ RNA of estradiol-stimulated male Oreochromis aureus was ligated to PTZ-19R and transformed into E. coli. TG-1 to yield 6 x 10(4) clones/ug DNA, of which 6% was vitellogenin positive. Restriction maps indicate 4 possible subgroups of vitellogenin cDNA, with homology, observed by Southern hybridisation, at a fragment flanked by Pst 1 sites. When the amino acid sequence derived from the DNA sequence of pOA Vg 62 was aligned with that of S. gairdneri, X. laevis, C. elegans and D. melanogaster, homologies of 63%, 22%, 21% and 15%, respectively, were obtained. Estradiol induced a vitellogenin gene transcript of 6500 nucleotides at 6 h and reached a maximum at 72 h after stimulation.

Amino Acid Sequence↗

Blood steroid levels in the goldfish: measurement of six ovarian steroids in small volumes of serum by reverse-phase high-performance liquid chromatography and radioimmunoassay.

A reliable and rapid technique for the measurement of estradiol-17 beta, testosterone, progesterone, 17 alpha-hydroxyprogesterone, 17 alpha,20 beta-dihydroxy-4-pregnen-3-one, and cortisol by reverse-phase high-performance liquid chromatography and radioimmunoassay in single female goldfish serum samples of 50 microliters was developed. The steroids were extracted with Sep-Pak C18 cartridges after heat treatment. While estradiol-17 beta was assayed directly after extraction, the other steroids were separated on a mu Bondapak C18 stainless-steel column with acetonitrile:water (53:47, v/v) in 17 min under isocratic conditions, and then quantitated by specific radioimmunoassays. The technique was validated and was shown to be highly accurate, precise, sensitive, and specific for measuring those particular ovarian steroids in goldfish serum. The steroid levels measured in this way were not significantly different from those measured after separation on a Nova-Pak C18 column with methanol:water (56:44, v/v), where all the individual steroids could be completely resolved during a 40-min run. With this technique, the steroid levels in the serum of the goldfish during the secondary yolk stage, the tertiary yolk stage, and at 0 hr after ovulation were determined. While estradiol-17 beta was found to be significantly higher in the tertiary yolk stage, testosterone was significantly higher in the secondary yolk stage than in the other two stages. There was no significant difference in the levels of cortisol, 17 alpha-hydroxyprogesterone,17 alpha,20 beta-dihydroxy-4-pregnen-3-one, and progesterone among the three stages. We conclude that the technique is particularly useful for assaying multiple steroids in very small volumes of biological fluids.

Animals↗