Ontogeny of thyroid hormone receptors in the brushtail possum (Trichosurus vulpecula)

Exercise electrocardiogram (ECG) has a high rate of false negative results in comparison with simultaneously performed thallium-201 perfusion scintigraphy, particularly in patients with single-vessel coronary artery disease, low exercise workload, inadequate heart rate rise, and resting ECG abnormalities. We present the case of a patient in whom thallium-201 SPECT scintigram revealed equally extensive and severe myocardial ischemia in two myocardial planes opposite each other. The accompanying exercise ECG did not disclose ischemic changes despite the adequacy of heart rate rise in this patient with severe right and left anterior descending coronary artery disease. We propose, as an explanation for this phenomenon, that in this patient the ischemic ST-segment vectors of equal magnitude and direction but of opposite sense, generated during stress, cancelled each other ("ischemic ST-segment counterpoise"), thus rendering the exercise ECG normal.     

A light microscopic study of oogenesis in the brushtail possum Trichosurus vulpecula

4-Methylquinoline (4-MeQ) showed an extraordinarily potent mutagenicity when compared to quinoline and isomeric methylquinolines. The major metabolite of 4-MeQ was 4-hydroxymethylquinoline, which was not mutagenic under the assay condition employed. Deuteration of the methyl group of 4-MeQ resulted in a decrease in the amount of the hydroxymethyl metabolite and an increase in mutagenicity, indicating that hydroxylation of the substituent methyl group is a detoxication process. A 3-chloro derivative of 4-MeQ was proven to be non-mutagenic. 4-Ethyl-quinoline, as well as 4-hydroxymethylquinoline, was much less mutagenic than 4-MeQ. Taking account of the structure-mutagenicity relationship, a possible mechanism is proposed for the potent mutagenic potential of 4-MeQ.     

Pathological studies of wobbly possum disease in New Zealand brushtail possums (Trichosurus vulpecula)

Following an outbreak of wobbly possum disease in a colony of brush tail possums (Trichosurus vulpecula), the disease was established experimentally in captive possums by inoculating the animals intraperitoneally with tissue homogenates. Crude tissue homogenates of liver remained infectious after freezing at -75 degrees C or filtration through a 0.22 micron filter. The disease was characterised by docility, incoordination, loss of balance and wasting. Fifteen of 16 infected animals had to be euthanased owing to the severity of clinical signs. Cachexia was the only change observed postmortem. Histology revealed widespread perivascular infiltrations with plasma cells and lymphocytes which were severe in the liver and kidney and moderate to mild in a variety of other tissues, including skeletal and cardiac muscle. Changes in the brain consisted of a mild to moderate mononuclear perivascular cuffing. Most of the animals had small to large numbers of circulating nucleated red blood cells and eosinopenia when they were euthanased. There was a consistent decrease in serum albumin concentration and an increase in serum globulins, which resulted in a decreased albumin:globulin ratio. Virus-like particles were observed in preparations of liver from two animals; they appeared to be spherical or icosahedral and were 45 nm in diameter.     

Purification and characterisation of hepatic glutathione transferases from a herbivorous marsupial, the brushtail possum (Trichosurus vulpecula)

In 1994, as part of their participation in the University of North Carolina Alumni Heart Study, 1101 women aged 45-51 years answered questions about their menopausal status and current use of hormone replacement therapy (HRT). Little is known about the use of HRT in younger women. We were interested in determining both patterns of HRT use and patient characteristics associated with HRT use in this cohort of women approaching the average age of menopause. After excluding women with breast, endometrial, and ovarian cancer, we studied 1080 women. These women identified themselves as: "There is no indication that I am near menopause" (stage 1, n = 326), "I think I may be close to or in the beginning stages of menopause but am not sure" (stage 2, n = 410), "I have begun menopause" (stage 3, n = 202), and "I have been through menopause" (stage 4, n = 142). The overall rate of HRT use was 22% (0% in stage 1, 8% in stage 2, 52% in stage 3, and 76% in stage 4). Both patterns of HRT use and patient characteristics associated with HRT use differed based on the woman's perception of her menopausal stage. In logistic regression models, where HRT use was the outcome variable, independent predictors of HRT use included stage of menopause, having had a hysterectomy, having had a bilateral oophorectomy, no family history of breast cancer, having had a pelvic examination in the last year, being married, and not participating regularly in physical exercise. A woman's perception of her stage in the process of reproductive aging correlates with her use of HRT. Informed decision making about HRT use should be tailored to the individual's perception of her menopausal stage.     

The Australian brushtail possum (Trichosurus vulpecula) gonadotrophin alpha-subunit: analysis of cDNA sequence and pattern of expression

Substantial evidence has accumulated to suggest that in the near future implementation of the veto-cell-suppressor concept in the treatment of kidney allograft recipients might lead to the establishment of life-long specific allograft tolerance in the absence of further immunosuppressive therapy. Veto suppression prevents the generation of antigen-specific T-helper and cytotoxic T lymphocytes in vitro provided that the T-lymphocyte precursors specifically recognize antigenic peptides associated with the major histocompatibility complex molecules class II and class I, respectively, expressed on the surface of the veto-active cell. Data from a large number of experimental and clinical studies strongly indicate that veto-active cells function in vivo and are capable of preventing allograft rejection. Thus, donor-cell-mediated veto activity is the most likely explanation for the well-known graft tolerizing effect of pretransplant donor blood transfusions in kidney graft recipients. A prerequisite for a veto-active environment in vivo is the establishment of lymphoid microchimerism, in which veto-active donor and recipient cells mutually downregulate potential alloaggression.     

cDNA sequence analysis, gene expression and protein localisation of the inhibin alpha-subunit of Australian brushtail possum (Trichosurus vulpecula)

A reliable source of human neural tissue would be of immense practical value to both neuroscientists and clinical neural transplantation trials. In this study, human precursor cells were isolated from the developing human cortex and, in the presence of both epidermal and fibroblast growth factor-2, grew in culture as sphere shaped clusters. Using traditional passaging techniques and culture mediums the rate of growth was extremely slow, and only a 12-fold expansion in total cell number could be achieved. However, when intact spheres were sectioned into quarters, rather than mechanically dissociated, cell cell contacts were maintained and cellular trauma minimised which permitted the rapid and continual growth of each individual quarter. Using this method we have achieved a 1.5 million-fold increase in precursor cell number over a period of less than 200 days. Upon differentiation by exposure to a substrate, cells migrated out from the spheres and formed a monolayer of astrocytes and neurons. No oligodendrocytes were found to develop from these human neural precursor cells at late passages when whole spheres were differentiated. This simple and novel culture method allows the rapid expansion of large numbers of non-transformed human neural precursor cells which may be of use in drug discovery, ex vivo gene therapy and clinical neural transplantation.     

Timing of zona pellucida formation in the tammar wallaby (Macropus eugenii) and brushtail possum (Trichosurus vulpecula)

The zona pellucida (ZP) is an extracellular coat that surrounds the mammalian egg, and serves as the primary recognition site for fertilizing spermatozoa. The timetable of ZP formation was examined in two marsupials, the tammar wallaby (Macropus eugenii) and the brushtail possum (Trichosurus vulpecula) using conventional histological methods, immunofluorescence and electron microscopy. Ovaries from tammar wallaby pouch young less than 80 days of age contained only primordial follicles with a single layer of flattened granulosa cells. There was no evidence of ZP formation until 98 days, when a small number of eggs surrounded by a single layer of cuboidal granulosa cells had a ZP detectable by periodic-acid-schiff staining and rabbit anti-pig ZP polyclonal antibody labelling. Possum ovaries at 108 and 114 days also contained a small number of eggs with a ZP and a single layer of cuboidal granulosa cells. The antibody also labelled the peripheral cytoplasm of oocytes at this stage and, occasionally, the granulosa cells. Antral follicles were first detected at 144 days in the wallaby and 125 days in the possum, and always contained an egg surrounded by a ZP. Ovaries from 147, 158, 165, 181, 184 and 210-day-old tammar wallabies contained a range of follicle types from primordial through early antrum formation. Electron microscopy confirmed observations made at the light microscope level. The ZP was first detectable in small primary follicles with a single layer of cuboidal granulosa cells in areas where microvilli had begun to form on the egg plasma membrane. Immunogold labelling indicated the egg cytoplasm as the origin of the ZP proteins. The ZP completely filled the space between the egg and the adjacent granulosa cells in preantral follicles, so that there was no perivitelline space.     

Physiological changes in brushtail possums, Trichosurus vulpecula, transferred from the wild to captivity

The goal of the present study was to examine the interaction of cholecystokinin (CCK-33) and its fragments: C-terminal octapeptide (CCK-8) and C-terminal tetrapeptide (CCK-4) with alpha- and beta-adrenoceptor agonists and antagonists. The effect of this interaction on arterial blood pressure and function of isolated heart was studied in rats. The results indicate that: 1) CCK-33 enhances the influence of catecholamines: noradrenaline and isoprenaline, mainly on the function of isolated heart. This peptide does not change cardiovascular effects of alpha-adrenoceptor antagonist--phentolamine. CCK-33 diminishes the influence of propranolol on the function of isolated heart. The hypotensive effect of beta-adrenoceptor antagonist is not affected by CCK-33. 2) CCK-8 does not alter cardiovascular effects of noradrenaline and isoprenaline. The peptide diminishes the hypotensive effect of phentolamine and reverses the hypotensive effect of propranolol. CCK-8 enhances the influence of propranolol and does not change the influence of phentolamine on the function of isolated heart. CCK-8 enhances bradycardia evoked by propranolol. 3) CCK-4 does not change the influence of noradrenaline and isoprenaline on arterial blood pressure and diminishes the hypotensive effect of phentolamine and propranolol. The peptide does not change cardiac effects of noradrenaline and diminishes the effects of isoprenaline, phentolamine and propranolol. On the basis of the present study, we concluded that CCK-33 and its fragments CCK-8 and CCK-4 modify the cardiovascular action of alpha- and beta-adrenoceptor agonists and antagonists. We suggest that effects we have observed correlate with the activation of the CCK-A receptors (CCK-33, CCK-8) or CCK-B receptors (CCK-4). CCK-related peptides may increase or reduce the effects of catecholamines indirectly through activation of alpha-adrenoceptors. We can not exclude direct action of the peptides on the heart.     

Morphology and histochemistry of the mandibular gland of the Australian brush-tail possum Trichosurus vulpecula (Marsupialia)

A histological, ultrastructural and histochemical study of the mandibular gland of the Australian possum Trichosurus vulpecula revealed essentially similar features as those described earlier for the mandibular gland of the taxonomically relatively unrelated American opossum Didelphis. The secretory endpieces consist of a branched tubular part, composed of serous cells whose secretion granules contain neutral glycoproteins, and terminal acini, consisting of seromucous cells containing small amounts of sialomucins. Relatively short intercalated ducts lead to striated ducts of variable ultrastructural appearance. The striated ducts run in bundles in the center of each sublobule of the gland. The possible functional significance of the abundance and variability in ultrastructure of the striated ducts is discussed.     

SmI cortical barrels in an Australian marsupial, Trichosurus vulpecula (brush-tailed possum): structural organization, patterned distribution, and somatotopic relationships

This study reports on the cerebral cortex of an Australian marsupial, Trichosurus vulpecula (brush-tailed possum). It consists of an analysis of layer IV of somatosensory cortex in tangential sections of flattened specimens and in oblique radial sections stained to show Nissl substance or myelin, or tested for succinic dehydrogenase. It includes results of electrophysiological mapping experiments that ascertained the somatotopic significance of the cytoarchitecture of this cortical region. Layer IV has two interlocking cytoarchitectural fields: one granular (the barrelfield, comprising cell-dense barrels 150 to 500 microns in diameter) and one dysgranular. Only neurons within the granular field responded to light cutaneous stimulation. In the barrelfield cell-sparse septa (about 100 microns wide), low in succinic dehydrogenase activity and containing many radial myelinated axons, separate adjoining barrels. Possum barrels are "solid," lacking the prominent hollows characteristic of most rodent barrels. In some specimens three to five small neuronal "lobules" may constitute each large barrel. In tangential sections the size, shape, and arrangement of barrels combine to form a histological caricature of the possum's body, especially of the face and forepaw. Six rows of "mystacial barrels" are homeomorphic to the six rows of large mystacial vibrissae, and "forepaw barrels" are homeomorphic to the glabrous palmar and apical digital pads. Correlating cortical recording sites and receptive fields confirmed that individual barrels represent specific cutaneous regions. These results show that the cortical barrels of brush-tailed possums are remarkably similar to those of rodents, in structure, arrangement, and functional significance.     

Isolation, characterization and radioimmunoassay of luteinizing hormone in the brushtail possum

Luteinizing hormone (LH) was purified from brushtail possum (Trichosurus vulpecula) pituitary glands. The purification procedure consisted of ammonium sulfate precipitation followed by triazinyl-dye chromatography, hydrophobic interaction chromatography and gel filtration. A yield of 10 microg LH g-1 pituitary with a recovery of 20% was obtained from 1400 pituitary glands (20.3 g). Contamination with possum follicle-stimulating hormone (FSH) was < or =0.05%. The amino acid analysis and the N-terminal sequencing for 10 cycles revealed close homology with LH from other mammals. Minor amounts of LH that had been truncated near the N-terminal were also detected. No contaminating proteins were found by amino acid sequencing. The potency of possum LH was 20% that of ovine LH in a receptor assay using possum testicular receptors and 4% that of ovine LH when bovine corpora lutea receptors were used. Possum LH was able to stimulate production of cyclic adenosine 3',5'-monophosphate by bovine granulosa cells. A radioimmunoassay (RIA) for possum LH using 125I-possum LH and an antiserum raised against ovine LH was developed. The RIA has a sensitivity of 0.15 ng mL-1, a 50% displacement of 1.9 ng mL-1 and a cross-reactivity of <0.02% against possum FSH. Plasma concentrations were 0.24+/-0.04 ng mL-1 (n = 8) and 0.39+/-0.12 ng mL-1 (n = 8) in female and male possums respectively. Administration of mammalian gonadotrophin-releasing hormone (GnRH) and chicken GnRH II stimulated increases in plasma LH concentrations in male and female possums. When comparing LH responses with administration of mammalian GnRH or chicken GnRH II, plasma LH concentrations remained elevated for a longer period of time in males than in females (P < 0.01); plasma LH concentrations also remained elevated for longer after mammalian GnRH than after chicken GnRH II (P < 0.01). Gonadectomy stimulated an increase in plasma concentrations of LH in both male (P < 0.01) and female (P < 0.05) possums. The rate of increase in plasma LH concentrations in males was faster than that in females. In summary, we have purified, partially characterized, and developed a RIA for possum LH.     

Lysine and glutamate transport in the erythrocytes of common brushtail possum, Tammar Wallaby and eastern grey, kangaroo

STUDY OBJECTIVE: To compare drug output from a vented nebulizer (Pari LC Jet Plus) with a traditional unvented nebulizer (Hudson 1730 T Up-Draft 11) using aerosolized tobramycin, which is frequently used in the treatment of cystic fibrosis. DESIGN: Six nebulizers of each type were filled with a 4 mL tobramycin (80 mg) solution and were driven by a compressor (Pulmo-Aide). Various inspiratory flows (VI) (0, 5, 10, 15, 20 L/min for the Pari LC Jet Plus and 0, 5, and 10 L/min for the Hudson 1730, all at 40% relative humidity) were directed through each nebulizer. Drug output was measured from changes in weight and concentration (assessed by changes in osmometry) within the nebulizer. Particle size distributions were determined by laser diffraction allowing the calculation of the amount of aerosol output in the respirable range (<5 microm). The nebulizers were first run until end-nebulization to establish total drug output and then for either 4 or 5 min to determine the rate of drug output (mg/min) before intermittent aerosol output. RESULTS: The total drug output without VI for both the unvented and the vented nebulizers was not significantly different, 55 (51, 60) mg for the Hudson 1730 vs 51 (49, 53) mg for the Pari LC Jet Plus (mean [95% confidence limits]). Inspiratory flow had no effect on the unvented Hudson 1730 nebulizer but significantly increased the rate of total drug output and the rate of drug output in the respirable range for the vented Pari LC Jet Plus nebulizer (VI=0, 3.35 [2.84, 3.85] and 1.72 [1.48, 1.96] compared with VI=20, 9.87 [9.03, 10.70] and 6.11 [5.33, 6.88] mg/min). CONCLUSIONS: These findings indicate that the increase in the rate of drug output with VI for the vented nebulizer would result in shorter nebulization times and a relative decrease in drug loss during the expiratory phase.     

Cytochrome P450 enzyme activities in the Australian brushtail possum trichosurus vulpesula: a comparison with the rat, rabbit, sheep and chicken

630 neonates with risk factors of perinatal hearing impairment were screened of hearing loss by means of registration of transient otoacoustic emissions before discharge from the newborn nursery. Neonates were screened additionally by means of brainstem evoked response audiometry, if they had bilateral negative emissions. 810 healthy neonates were screened as control group. The investigations were carried out in incubator after the feeding of neonates. The prevalence of a bilateral negative cochlear response was 5.2% in the risk babies and 1.7% in healthy neonates. Neonates are high risk patients for hearing loss if they show craniofacial anomalies including alcohol embryofetopathy, connatal infections, or very low birth weight babies with additional risk factors. The pedaudiological control investigations of the babies with a negative bilateral cochlear response delivered in the risk group 15 cases (2.4%) with an important hearing impairment and in the healthy neonates 2 cases (0.25%) respectively. Prevalence and importance of perinatal hearing impairment explains the necessity of detection in the neonatal period.     

A role for thyroid hormones in the regulation of diet-induced thermogenesis in birds

The possible involvement of thyroid hormones in avian diet-induced thermogenesis (DIT) was investigated in two lines of cockerels divergently selected for high (R-) or low (R+) food efficiency. For a given body weight, R+ cockerels exhibited a higher food intake than R- cockerels (+49 to +76%) and increased DIT (+25%). Plasma thyroxine (T4) level did not differ between lines whatever the feeding status of the birds. Plasma 3,5,3'-triiodothyronine (T3) level was lower in fasted R+ than in fasted R- cockerels while the opposite was observed after a meal. Iopanic acid injections reduced both plasma T3 concentrations and heat production to the same levels in both lines. Hepatic 5'-deiodinase activity measured with an exogenous sulfhydryl group (dithiothreitol) did not differ between lines, but when the sulfhydryl group was omitted, the activity was higher in R+ than in R- birds (90 v. 42 pmol T3/min per liver). T3-binding capacity of isolated hepatic nuclei was higher (+76%) in R+ than in R- birds. Long-term or acute pair-feeding of R+ cockerels to the level of R- controls did not alter these results. The present results suggest that T3, mainly originating from peripheral conversion of T4 to T3, is involved in DIT in the R+ line. Availability of endogenous sulfhydryl groups appears to play an important part in the modulation of hepatic deiodinase activity. The higher concentration of nuclear T3 receptors may further increase the effects of the hormone, suggesting a major role of thyroid hormones associated with catecholamines in the stimulation of avian DIT. The underlying thermogenic mechanisms remain to be elucidated.     

The role of ultrasonography in thyroid disease

Ultrasound examination of the thyroid gland is widely used in the diagnosis of thyroid disease. This test is easy and rapid to perform, widely available and the results are readily interpreted. Using ultrasound the image of foci of disease within the gland are easily identified, especially using high frequency probes which enable solid nodules up to approx 3 mm to be revealed with 10 MHz probes. In non-nodular thyroid disease the ultrasonic structure guides the diagnosis (thyroiditis, Graves' disease). During the follow-up of thyroidectomised patients ultrasound can easily reveal postoperative anatomic variations and an early diagnosis can be obtained of any signs of local recurrence of the primary disease. Only hemiagenesia and hypoplasia can be accurately evaluated in congenital disease, whereas in the event of the persistence of the thyroglossal duct the latter can only be diagnosed if it presents a cystic evolution. Thyroid ectopia cannot be identified and must be studied using thyroscintigraphy, preferably performed using 131I as the isotope. The acquired pathology is classified into phlogistic processes, diffuse or nodular hyperplasia, benign and malignant neoplasia. This classification is widely accepted by virtually all authors. In thyroiditis, ultrasound may facilitate the diagnosis of De Quervain's non-suppurative sub-acute thyroiditis (TANS) and Hashimoto's chronic thyroiditis, although always in association with clinical and laboratory tests. The most frequent thyroid pathology is without doubt goitre. This disorder may occur in a non-nodular (widespread goitre with an endemic or sporadic pattern) or nodular form which may be single or multiple. The term goitre is used to indicate the increased volume of the thyroid gland independently of the causes which have provoked it. Common goitre is defined as being endemic when in some geographic area 10% of the general population or 20% of the school-aged population suffers from thyroid hyperplasia (areas of goitrogenic endemic disease). Graves' disease may be included in the group of thyroid hyperplasia diseases, although it is distinguished from the simple versions by the marked glandular hyperactivity which creates manifest hyperthyroidism. In this pathology ultrasonography must be supplemented by colour-Doppler wherever possible. Thyroid nodules are subdivided in terms of their echostructure into 5 types: liquid, mixed (prevalently solid or prevalently liquid), hyperechogenic solid, isoechogenic solid and hypoechogenic solid. The characteristics of benign nodules are: hypoechogenic structure, regular edges, complete and uniform hypoechogenic peripheral halo.(ABSTRACT TRUNCATED AT 400 WORDS)     

Role of thyroid hormones in early postnatal development of skeletal muscle and its implications for undernutrition

Energy intake profoundly influences many endocrine axes which in turn play a central role in development. The specific influence of a short period of mild hypothyroidism, similar to that induced by undernutrition, in regulating muscle development has been assessed in a large mammal during early postnatal life. Hypothyroidism was induced by providing methimazole and iopanoic acid in the feed of piglets between 4 and 14 d of age, and controls were pair-fed to the energy intake of their hypothyroid littermates. Thyroid status was evaluated, and myofibre differentiation and cation pump concentrations were then assessed in the following functionally distinct muscles: longissimus dorsi (l. dorsi), soleus and rhomboideus. Reductions in plasma concentrations of thyroxine (T4; 32%, P < 0.01), triiodothyronine (T3; 48%, P < 0.001), free T3 (58%, P < 0.001) and hepatic 5'-monodeiodinase (EC 1.11.1.8) activity (74%, P < 0.001) occurred with treatment. Small, although significant, increases in the proportion of type I slow-twitch oxidative fibres occurred with mild hypothyroidism, in l. dorsi (2%, P < 0.01) and soleus (7%, P < 0.01). Nuclear T3-receptor concentration in l. dorsi of hypothyroid animals compared with controls increased by 46% (P < 0.001), a response that may represent a homeostatic mechanism making muscle more sensitive to low levels of circulating thyroid hormones. Nevertheless, Na+, K(+)-ATPase (EC 3.6.1.37) concentration was reduced by 15-16% in all muscles (l. dorsi P < 0.05, soleus P < 0.001, rhomboideus P < 0.05), and Ca(2+)-ATPase (EC 3.6.1.38) concentration was significantly reduced in the two slow-twitch muscles: by 22% in rhomboideus (P < 0.001) and 23% in soleus (P < 0.05). It is concluded that during early postnatal development of large mammals a period of mild hypothyroidism, comparable with that found during undernutrition, induces changes in myofibre differentiation and a down-regulation of cation pumps in skeletal muscle. Such changes would result in slowness of movement and muscle weakness, and also reduce ATP hydrolysis with a concomitant improvement in energetic efficiency.