Search PubMed⌕ Search

Biomedical subjects

D M Robertson

Publications and source records attributed to D M Robertson.

At least 235 records · Page 13Linked to original sources

Isolation of inhibin from bovine follicular fluid.

Bovine follicular fluid was used as a source for the isolation of gonadal inhibin, the activity of which was monitored by the dose dependent suppression of the FSH content of cultured pituitary cells. The procedures presented result in over 3000-fold purification of the starting material and the purified inhibin has an apparent molecular weight of 56000. The purified inhibin can be dissociated under reducing conditions into two subunits with molecular weights of 44000 and 14000 daltons.

Animals↗

Neurotoxicity of Adriamycin and misonidazole in the mouse.

The neurotoxicity of the anticancer drug adriamycin was investigated in the peripheral nerve of the mouse. Injection of adriamycin into the sciatic nerve resulted in biochemical and morphologic signs of severe axonal degeneration. The biochemical evidence was based on marked increases in lysosomal enzyme activity. Acid protease and N-acetylglucosaminidase remained elevated for more than 70 days after injecting the drug. In contrast, local injection of the radiation-sensitizing drug misonidazole into the peripheral nerve failed to increase lysosomal enzyme activity.

Animals↗

Electrofocusing fractionation of follicle stimulating hormone in pituitary cell culture extracts from male and female rats.

Three-day pituitary cell cultures from adult male and female rats were incubated for 4 h in the presence of 10 nM LHRH and the molecular heterogeneity of FSH was assessed in the media of LHRH-stimulated cells and in cell extracts from unstimulated cells using an electrofocusing technique. The pI distribution of FSH showed a high degree of similarity between cell media and cell extracts of each sex although differences were observed between sexes. Pituitary cell cultures from male rats were also incubated in the presence of 10(-8) M testosterone and 10(-8) M estradiol and the pI distribution of FSH from media after LHRH stimulation was determined. No significant differences in the pI profiles were observed. Incubation with charcoal-treated bovine follicular fluid (an inhibin source) resulted in a significant reduction in recovered FSH activity in the pH region 3.61-3.92 although this decrease did not markedly alter the pI profile of FSH. Close similarities were observed in the pI distribution of FSH of pituitary cell culture extracts and pituitary gland extracts from intact animals of both sexes, however, differences in pI distribution were noted in pituitary extracts in the male but not the female following gonadectomy. It is concluded that (1) stored FSH is released from the pituitary without major modification to its structure as assessed from its pI profile, (2) sex differences in the pI profile of FSH in pituitary extracts are retained in culture and following LHRH-stimulated release, (3) the pI distribution of FSH is not affected by testosterone or estradiol and only minimally by inhibin in short-term cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Inhibin: 1985 update on action and purification.

Inhibin is a gonadal hormone, which exerts a specific negative feedback action on the pituitary secretion of follitropin (FSH) in male and female animals. The existence of inhibin was postulated over 60 years ago. Yet, until recently, little progress has been made in its isolation and characterisation. This lack of progress may be attributed to several factors: first, the use of a variety of assay systems of ill-defined specificity, secondly, the use of a variety of sources for unpurified inhibin, and thirdly, the inability of investigators to purify inhibin using classical purification procedures. During the last few years, and in particular during the last year, several publications on the isolation and characterisation of inhibin have appeared. This review attempts to place the various reports on the nature of inhibin into perspective. It is concluded that there are at least 2 classes of proteins with inhibin-like activity: a relatively large molecular weight material with apparent molecular mass between 40 and 70 kDa found in gonadal extracts and fluids, and a smaller material, with molecular mass between 5 and 20 kDa, found in seminal plasma. However, the observations that various purified seminal plasma inhibin preparations are either inactive in in vitro assays used to characterize gonadal inhibin or have been shown to be prostatic in origin suggest that they are unlikely to be involved in the gonadal regulation of FSH secretion. It has yet to be established if the purified gonadal inhibin preparations are the biological active forms involved in controlling FSH secretion in vivo.

Animals↗

The isolation and measurement of luteinizing hormone-releasing hormone (LHRH) from the rat testis.

Although LHRH has been implicated in the direct control of rat Leydig cell function, LHRH has not been previously detected in the rat testis. An optimized fractionation procedure, which involved acid extraction, bulk fractionation on ODS-silica, extraction in chloroform/ethanol, ether extraction, gel filtration on Sephadex G-15 and RP-HPLC, was employed to isolate LHRH from lyophilized adult rat testes. LHRH activity was assessed by an in vitro LHRH bioassay system employing rat anterior pituitary cells in monolayer culture and several radioimmunoassays for LHRH. LHRH recoveries were monitored by the addition of exogenous LHRH to some samples of lyophilized testes prior to extraction. LHRH bioactivity was non-detectable in the LHRH region of the gel filtration profile of testis extracts in the absence of exogenous LHRH; however, using the most specific LHRH RIA procedure, LHRH immunoactivity which co-chromatographed with LHRH following RP-HPLC was found in all extracts. Based on an average LHRH recovery of 31.0%, as determined from the extracts containing exogenous LHRH, the level of endogenous LHRH immunoactivity in the extracts was determined to be equivalent to 5.6 pg LHRH/g dry weight or 1.0 pg LHRH/testis. These results indicate that the levels of LHRH in the adult rat testis are considerably less than those of the hypothalamus. Based on these findings a simplified fractionation/assay procedure with sufficient sensitivity can be devised for the quantitation of testicular LHRH as a means of clarifying its physiological role in gonadal function.

Animals↗

Morphological and functional characterization of interstitial cells from mouse testes fractionated on Percoll density gradients.

Cell fractions were obtained by separation on Percoll density gradients after dissociation of mature mouse testes by mechanical or collagenase dispersion, and the ultrastructure, hCG receptor properties, and hCG-stimulated testosterone (T) production of these fractions were compared. Gradients were fractionated according to specific gravity, and all cell types were quantitated using morphometric techniques. Three peaks of specific [125I]iodo-hCG binding corresponding to densities of 1.0667 g/cm3 (fractions 2-3), 1.045 g/cm3 (fractions 6-7), and 1.0365-1.0215 g/cm3 (fraction 9) were obtained after collagenase dispersion, but the second peak of binding (fractions 6-7) was not observed after mechanical dispersion. Morphometric studies were performed by light microscopy on the cells present in the fractions corresponding to the first and second hCG binding peaks obtained by either mechanical or collagenase dispersion; the third representing germ cells and membranous debris was not studied further. Regardless of the method of preparation, morphologically intact Leydig cells represented 60-80% and 7-10% of the cells in fractions 2-3 and 6-7 that were associated with the first and second peaks of hCG binding Leydig cells obtained from fractions 6-7 contained greater numbers of lipid inclusions than Leydig cells from fractions 2-3. Mechanically dispersed Leydig cells exhibited similar numbers of hCG receptors in the dense and light Leydig cells, but hCG stimulated T production per Leydig cell was significantly greater in the dense Leydig cells containing few lipid inclusions. T production by dense and light collagenase-dispersed Leydig cells was not significantly different. The second hCG binding peak in collagenase-dispersed cell fractions 6-7 was associated with an increase in the number of an indeterminate connective cell type released from the testes by collagenase treatment, whose ultrastructure and limited hCG-binding capacity suggested that they may represent Leydig cell precursors. It is concluded that the identification and quantitation of different cell types in isolated testicular cell fractions is, therefore, of fundamental importance in the interpretation of receptor and secretory capacities of enriched preparations of Leydig cells.

Animals↗

Effects of hypophysectomy and subsequent FSH and testosterone treatment on inhibin production by adult rat testes.

The hormonal control of inhibin production by adult rat testes was investigated using an in-vitro inhibin bioassay validated for the measurement of inhibin activity in charcoal-treated rat testicular extracts. The effect of hypophysectomy examined at 16 h, 3, 7 and 42 days after surgery showed a decrease in testicular inhibin content and seminiferous tubule fluid production by 7 days and a decrease in inhibin production by 42 days. Serum FSH and LH were suppressed 3 days after surgery. In 30-day chronically hypophysectomized adult rats treated for 3 days with twice daily s.c. injections of (a) human FSH (hFSH, 22 i.u./rat per day), (b) testosterone (5 mg/rat per day), (c) hFSH + testosterone (same doses as a and b), or (d) human chorionic gonadotrophin (hCG, 12 i.u./rat per day), hFSH or hFSH and testosterone stimulated an increase in testicular inhibin content but not in inhibin production or tubule fluid production. Testosterone and hCG had no effect on these parameters. It is concluded that in vivo, FSH alone stimulates an increase in testicular inhibin content. The failure to observe an increase in inhibin production in vivo is attributed to the suppression of seminiferous tubule fluid production under the same experimental conditions.

Animals↗

Polyneuropathy with vagus and phrenic nerve involvement in breast cancer. Report of a case with spontaneous remission.

Dysphagia, respiratory insufficiency, and bilateral vocal cord paralysis developed in a 70-year-old woman seven years after resection of a breast carcinoma. There was spontaneous partial remission of symptoms before death, in the absence of treatment. Necropsy showed both inflammatory and metastatic infiltrates of vagus nerves, with demyelination out of proportion to axonal loss. Spontaneous resolution of metastases-evoked inflammation within vagus and phrenic nerves may have been the basis of clinical remission.

Aged↗

Chlorambucil in the treatment of uveitis and meningoencephalitis of Behçet's disease.

Among 21 patients with Behçet's disease, the results of various treatments were assessed. Treatment with chlorambucil, 0.1 mg/kg daily, compared favorably with corticosteroids. Uveitis and visual acuities improved in five of seven eyes when the patients were treated with chlorambucil, whereas improvement was observed in only four of 13 eyes when treatment consisted of corticosteroids. In 19 treatment courses among 14 patients with meningoencephalitis or recurrent meningitis, eight of nine patients treated with chlorambucil had remission of their disease; eight of 10 treatment courses with other agents--generally, corticosteroids--failed to induce remission in any of eight patients. Elevated cerebrospinal fluid cell counts returned to normal in five of nine patients treated with chlorambucil but remained elevated in five of six patients treated with corticosteroids. Two patients died, one who was taking prednisone plus cyclophosphamide and the other after prednisone plus multiple immunosuppressive treatments. Toxicity from chlorambucil included leukopenia (two patients), thrombocytopenia (one), bronchopneumonia (one), and amenorrhea (two). The duration of chlorambucil therapy averaged 1.8 years both for patients with uveitis and for those with meningoencephalitis.

Adolescent↗

Ophthalmic features of necrobiotic xanthogranuloma with paraproteinemia.

Necrobiotic xanthogranuloma of the skin is associated with paraproteinemia and, often, with plasma proliferative disorders, including multiple myeloma. Other commonly recognized systemic abnormalities include hepatosplenomegaly, a highly increased erythrocyte sedimentation rate, and leukopenia. Fifteen of 16 patients (seven men and nine women with a mean age of 54 years) with this condition had ophthalmic manifestations. Thirteen patients had lesions affecting the skin of the eyelids and periorbital tissue; on casual examination these lesions resembled plane xanthoma. Unlike plane xanthoma, however, the lesions of necrobiotic xanthogranuloma were almost always indurated. Further, the lesions frequently became inflamed, leading to superficial ulceration. Deeper lesions occasionally involved the orbit. Yellow lesions were sometimes visible in the episcleral tissues where they were associated with recurrent symptoms of scleritis and episcleritis. Biopsy specimens of the skin lesions disclosed a distinctive pattern of subepidermal granulomatous masses with focal aggregates of histiocytes and giant cells surrounded by hyaline necrobiosis. Surgical excision of the eyelid lesions was often followed by recurrence and increased activity of the lesions. Low-dose chemotherapy is likely to produce a favorable response, with regard to both the skin lesions and the paraprotein abnormalities.

Adult↗

A study of HLA antigens in alcoholism.

The concept that alcoholism may be inherited has been suggested on the basis of twin and adoption studies and the further evidence that alcoholism is commonly seen in several members of a family. Genetic marker studies that have been conducted among alcoholics have often yielded seemingly contradictory results. Thus far, genetic marker studies involving HLA antigens have not been used in the study of alcoholism. In the current study, we identified and performed HLA typing in 52 alcoholics to determine whether a significant difference existed between the frequency of HLA-A and HLA-B locus antigens in an alcoholic population and that in a control population consisting of 1,704 nonalcoholic blood donors. In this study, we could not demonstrate a difference in the occurrence of these antigens between the two populations.

Alcoholism↗

von Hippel-Lindau disease: a familial, often lethal, multi-system phakomatosis.

A review of 36 cases of von Hippel-Lindau disease underscores the important role of the ophthalmologist in evaluating this phakomatosis. Retinal angiomatosis occurred in almost two-thirds of those affected, was usually diagnosed before other target organs, and frequently led to decreased visual acuity. Common non-ophthalmic manifestations included cerebellar (69%), medullary (11%) and spinal (8%) hemangioblastoma, renal cell carcinoma (22%), and polycythemia (17%). Pheochromocytoma was a les common association. The most common causes of death were cerebellar hemangioblastoma and renal cell carcinoma. For the surveillance of suspected von Hippel-Lindau victims, a detailed family history and retinal examination is recommended. Essential laboratory tests include a complete blood count and computed tomography of the head, upper cervical region, and abdomen. Periodic pre-symptomatic screening is imperative for follow-up investigation of affected individuals to determine possible additional occult manifestations and of non-affected family members at risk to von Hippel-Lindau disease.

Adenocarcinoma↗

Pharmacological modification of blood-brain barrier permeability following a cold lesion.

The effect of desipramine, imidazole, thioridazine and trifluoperazine on blood-brain barrier (BBB) permeability after a 24 hour cold lesion was studied in rats. Changes in BBB permeability were determined using a quantitative horseradish peroxidase (HRP) assay. The four drugs tested did not alter the quantity of HRP in the cortex of control animals, or in the contralateral cortex of test animals. However, imidazole, desipramine and trifluoperazine significantly reduced the HRP extravasation in and around the cold lesion. Several mechanisms for this effect are suggested; one possible mechanism common to all these drugs is the reduction of increased vesicular transport in cortical vessels adjacent to the cold lesions.

Animals↗

Pituitary gonadotrophins in Booroola and control Merino sheep.

Pituitary content of FSH and LH, using radioreceptor assay methods, was determined in control and Booroola Merino ewes on the 3rd day of the oestrous cycle and in adult rams slaughtered in winter. Significantly more pituitary FSH (as per gland or per g wet wt) was found in the Booroola than in the control ewe. No significant differences were found in LH content although the difference in FSH/LH ratio between Booroola and control ewes was significant (P less than 0.001). Pituitary FSH content was similar in the rams of the two genotypes. A good correspondence between FSH values by the radioreceptor assay and by radioimmunoassays using anti-ovine and anti-human serum was observed for the Booroola ewes. However, radioimmunological estimates of FSH activity were significantly higher than radioreceptor estimates in control ewes and in Booroola and control rams in which the pituitary FSH values were 5-6% of that of the ewe. This over-estimation is attributed to differences in specificity between methods. Fractionation of pituitary extracts by electrofocussing indicated a similar pI profile of FSH for ewes and rams of both genotypes. It is concluded that quantitative rather than qualitative differences in pituitary FSH occur in Booroola and control Merino ewes. It is suggested that increased FSH levels contribute to the hormonal basis for the increased ovulation rate of the Booroola Merino.

Animals↗

An in-vivo method for estimating inhibin production by adult rat testes.

The concentrations of inhibin in samples of rat testicular venous and arterial blood and interstitial fluid were measured by an in-vitro bioassay using pituitary cells in culture in which the standard was an ovine testicular lymph preparation (assigned potency 1 unit/mg). Inhibin levels were undetectable (less than 2 U/ml) in both blood samples but reached a mean concentration of 120 +/- 7 U/ml in testicular interstitial fluid. After unilateral efferent duct ligation the rate of inhibin accumulation in seminiferous tubules was determined by the difference in the inhibin content of the ligated and unligated testes. Additionally, the rate of seminiferous tubule fluid production was obtained from the difference in weight between the ligated and non-ligated testes. In the 24 h after efferent duct ligation there were linear increases in inhibin (18.5 +/- 1.0 U/h) and in seminiferous tubule fluid production (26 +/- 1 microliter/h), but there were no changes in serum FSH and LH levels. Experimental induction of bilateral cryptorchidism led to a decrease in the inhibin content of the testis after 10 days. The rate of inhibin accumulation after efferent duct ligation declined more rapidly than the inhibin content, being significantly depressed in cryptorchid testes after 3 days, suggesting that this measurement is a more sensitive index of inhibin production than the determination of testicular inhibin content.

Animals↗

Relationship between testicular inhibin content and serum FSH concentrations in rats after bilateral efferent duct ligation.

The testicular inhibin content showed an initial increase in the first 2-3 days after bilateral ligation of the efferent ducts of rats, followed by a subsequent decline to levels significantly below normal by 14 days, and reached 25% of control values at 42 days. Serum concentrations of FSH and LH were significantly increased at Day 6-7 after treatment and were still elevated after 42 days. The decline in testicular inhibin content at times associated with elevated FSH concentrations is consistent with the hypothesis of inhibin being involved in the feedback control of FSH secretion.

Animals↗