PubMed Health⌕ Search

Biomedical subjects

S J Newsholme

Publications and source records attributed to S J Newsholme.

At least 19 recordsLinked to original sources
PubMed2000

Two-dimensional electrophoresis of liver proteins: characterization of a drug-induced hepatomegaly in rats.

Two-dimensional electrophoresis (2-DE) of liver proteins was applied to further characterize an unusual drug-induced increase in hepatocellular rough endoplasmic reticulum (RER) in Sprague-Dawley rats given a substituted pyrimidine derivative. Absolute liver weights of drug-treated rats (9.9 +/- 0.4 g) increased above vehicle-treated controls (7.2 +/- 0.2 g) by 37%. Light microscopy revealed diffuse granular basophilia of the hepatocellular cytoplasm, uncharacteristic of hepatocytes and suggested cells rich in ribosomes, which was confirmed by electron microscopy. Immunostaining for cell proliferation, viz., 5-bromo-2'-deoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA), indicated marked hepatocellular proliferative activity. 2-DE of solubilized liver using an ISO-DALT gel system indicated significant (p<0.001) quantitative changes in at least 17 liver proteins (12 increased, 5 decreased) compared to controls. The protein with the largest increase was homologous to acute-phase reactant, contrapsin-like protein inhibitor-6. Other markedly upregulated proteins were methionine adenosyltransferase, a catalyst in methionine/ATP metabolism and mitochondrial HMG-CoA synthase, involved in cholesterol synthesis. The complementary strategies of 2-DE coupled either with database spot mapping or protein isolation and amino acid sequencing successfully identified a subset of proteins from xenobiotic-damaged rodent livers, the expression of which differed from controls. However, the current bioinformatics platform for rodent hepatic proteins and limited knowledge of specific protein functionality restricted application of this proteomics profile to further define a mechanistic basis for this unusual hepatotoxicity.

Animals↗
PubMed1998

Intralesional cytokines in chronic oxazolone-induced contact sensitivity suggest roles for tumor necrosis factor alpha and interleukin-4.

An analysis was conducted of the cytokine profile and inflammatory response in oxazolone sensitized mouse skin. Following exposure to oxazolone, the intralesional production of inflammatory cytokines was demonstrable at the levels of both mRNA and protein. An initial challenge led to a transient increase in tumor necrosis factor-alpha production followed predominately by the T helper (Th)1 cytokine, interferon-gamma. There was a minimal production of interleukin-4, a Th2 cytokine. Continued exposure to oxazolone led to a downregulation of interferon-gamma and an upregulation of interleukin-4 production. A strong relationship was found between interleukin-4 and the inflammatory response, as measured by ear thickness. Similar experiments conducted in mast cell-deficient mice revealed reduced neutrophil influx but only minor changes in cytokine profile. An irritant response induced by chronic exposure of mouse skin to phorbol ester did not reveal any significant interferon-gamma or interleukin-4 response but was characterized by a tumor necrosis factor-alpha response that correlated with the inflammatory response. These observations suggest that the major source of interferon-gamma and interleukin-4 in the oxazolone response may be the infiltrating lymphocytes; whereas the tumor necrosis factor-alpha may result from the local irritation seen with both oxazolone and phorbol ester. At the end of 4 wk of chronic exposure to oxazolone, it was found that serum IgE levels had significantly increased. Histologic analysis of the skin lesion revealed that a mixed infiltrate including eosinophils developed upon repeat exposure to oxazolone. These findings are consistent with an early predominate Th1 response that is reduced and largely replaced with a Th2 response upon chronic T cell activation.

Animals↗
PubMed1996

Persistent airway eosinophilia after leukotriene (LT) D4 administration in the guinea pig: modulation by the LTD4 receptor antagonist, pranlukast, or an interleukin-5 monoclonal antibody.

Aerosolized cysteinyl leukotrienes (CysLTs) elicit migration of eosinophils into guinea pig lungs and the airways of patients with asthma. The present studies were designed to analyze the concentration-response relationship, time course, and pharmacologic and histologic characteristics of leukotriene D4 (LTD4)-induced eosinophil influx into the airways of conscious guinea pigs. Animals were exposed to aerosols of 0.3 to 30 microg/ml LTD4 for 1 min, during which specific airway conductance (sGaw) was monitored. Bronchoalveolar lavages (BALs) of guinea pig airways were conducted at selected times from 4 h to 4 wk after LTD4 challenge. LTD4 produced maximal decreases in sGaw (70 to 90% reduction) at all concentrations tested and concentration-related increases in eosinophil levels in BALs, assessed 24 h after challenge. Increased numbers of eosinophils in the bronchial epithelium and subepithelium were confirmed histologically. Significant eosinophilia was maintained for up to 4 wk postchallenge. Pretreatment with the LTD4 receptor antagonist, pranlukast (ONO-1078, SB 205312) (20 mg/kg, intragastrically), significantly inhibited both the bronchoconstriction and the eosinophilia at 24 h, whereas the cyclooxygenase inhibitor, meclofenamic acid (5 mg/kg, intragastrically), had no effect on either parameter. Histologic observations were consistent with BAL results. Pretreatment with the rat anti-mouse antibody to interleukin-5 (IL-5), TRFK-5 (10-300 microg, intraperitoneally), produced dose-related inhibition of LTD4-induced eosinophilia, measured in 24 h or 3 wk BAL, but did not affect the acute bronchoconstriction. These results indicate that LTD4 elicits airway eosinophil influx in guinea pigs which persists as long as 4 wk after a single exposure, and provide the first evidence that IL-5 may have a role in LTD4-induced airways inflammation. This and other previously reported proinflammatory effects of LTD4 may contribute significantly to its overall influential role in the pathophysiology of asthma, and may underlie the therapeutic benefit of CysLT receptor antagonists, such as pranlukast, in this disorder.

Animals↗
PubMed1994

Conjunctival leukocyte infiltration evoked by leukotrienes: differing responses among rodent species.

Application of leukotrienes (LT) B4 or D4 to the guinea-pig eye evokes conjunctival eosinophil infiltration, providing an in vivo model of tissue eosinophilia. To determine if other rodent species would respond similarly, LTB4 or LTD4 were applied to the right eye of male mice and rats (three per group) at doses of 250, 1000 or 5000 ng/eye (mice) and 1000 or 5000 ng/eye (rats). Left eyes received the same volume (mice, 5 microliters; rats, 20 microliters) of vehicle (isotonic saline). Conjunctivae were evaluated histologically 6 h after application. In mice, 1000 or 5000 ng/eye LTB4 resulted in mild conjunctival neutrophilia, but no leukocyte response to LTD4 was observed. In rats, no leukocyte responses, either to LTB4 or to LTD4, were observed. The results illustrate interspecies differences in leukocyte responsiveness to LT and indicate specifically that, unlike the guinea pig, neither mouse nor rat responds to LT with conjunctival eosinophilia.

Animals↗
PubMed1993

cAMP-specific phosphodiesterase inhibitor, rolipram, reduces eosinophil infiltration evoked by leukotrienes or by histamine in guinea pig conjunctiva.

The effect of rolipram, an isozyme IV-selective inhibitor of cAMP-specific phosphodiesterase, was evaluated in a guinea pig eye model of tissue eosinophilia. (R)-rolipram was administered by gavage to guinea pigs 1 h prior to topical ocular challenge with a mixture of leukotrienes (LTs) (10 ng LTB4 + 1000 ng LTD4/eye) or with histamine dihydrochloride (1 mg/eye). Conjunctivae were evaluated histologically 6 h after challenge. Eosinophil counts per millimeter of conjunctival epithelium in LT-challenged animals that received (R)-rolipram at dosages of 0.1, 0.3, 1, 3, or 10 mg/kg were reduced by 63, 63, 84, 81 and 90% respectively, compared to LT-challenged controls. Reduction was statistically significant (P < 0.05) at all dosages. Eosinophil counts per millimeter of epithelium in histamine-challenged animals that received 10 mg/kg (R)-rolipram were reduced by 79% compared to histamine-challenged controls (P < 0.01). The results indicate that (R)-rolipram inhibits the response to two distinct classes of mediator in this model of eosinophil infiltration, adding support to the contention that isozyme IV-selective cAMP phosphodiesterase inhibitors offer therapeutic potential for human asthma.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1993

Inhibition of antigen-induced bronchoconstriction and eosinophil infiltration in the guinea pig by the cyclic AMP-specific phosphodiesterase inhibitor, rolipram.

Selective inhibition of the low Km cyclic AMP-specific phosphodiesterase has been shown to inhibit inflammatory cell function and relax airway smooth muscle. These studies were conducted to characterize the bronchodilator and anti-inflammatory activity of rolipram, an archetypical cyclic AMP-specific phosphodiesterase inhibitor, in in vitro and in vivo guinea pig airway models. In isolated tracheal rings from ovalbumin (OA)-sensitive guinea pigs, both R- and S-enantiomers of rolipram (1 microM) significantly antagonized OA-induced contractions. In contrast, neither enantiomer at concentrations up to 1 microM significantly inhibited histamine- or LTD4-induced contractions. In superfusion and mediator release experiments, both enantiomers of rolipram significantly reduced antigen-induced prostaglandin D2 release, but had minimal effect on histamine release. In anesthetized, ventilated OA-sensitive guinea pigs, racemic rolipram or enantiomers reduced OA-induced bronchoconstriction with ID50 values of approximately 0.25 mg/kg i.v. Histamine- and leukotriene D4-induced bronchoconstriction were not affected by doses of rolipram which abolished the response to OA. Higher doses (3-10 mg/kg) reduced histamine-, but not the leukotriene D4-induced bronchoconstriction. In conscious OA-sensitive guinea pigs, intragastric pretreatment with rolipram dose-dependently reduced both the OA-induced decreases in specific conductance as well as the corresponding pulmonary eosinophil influx as assessed by both bronchoalveolar lavage and histological evaluation. Therefore, rolipram produces significant inhibition of antigen-induced bronchoconstrictor and inflammatory responses, thus providing strong evidence that this pharmacological approach may be of significant therapeutic value in allergic asthma.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia↗
PubMed1992

Agonist-related differences in the relationship between cAMP content and protein kinase activity in canine trachealis.

We investigated the relationships between relaxation, cyclic AMP (cAMP) accumulation and cAMP-dependent protein kinase (cAMP-PK) activity in canine tracheal smooth muscle. In time course and concentration-response studies, forskolin and isoproterenol elicited relaxation of isolated trachealis strips that was accompanied by an increase in cAMP content and an activation of cAMP-PK. Although these results were consistent with the proposal that cAMP is a second messenger mediating relaxation of airway smooth muscle, close inspection of the data revealed a discrepancy in the relationship between cAMP accumulation and relaxation. To induce equivalent degrees of tracheal relaxation, forskolin generated greater increments in cAMP accumulation than did isoproterenol. On the other hand, the activation state of cAMP-PK correlated reasonably well with relaxation regardless of which agonist was used. Further analysis of the data revealed that the apparent disparity between cAMP accumulation and relaxation could largely be explained at the level of the relationship between cAMP content and cAMP-PK activity: compared to isoproterenol, forskolin induced greater increases in cAMP accumulation to achieve the same activation state of cAMP-PK. These observations lend support to the proposal that in canine trachealis, various components of the cAMP/cAMP-PK cascade exist in distinct subcellular compartments such that not all of the cAMP generated in response to forskolin has access to its target enzyme, cAMP-PK.

Animals↗
PubMed1988

Technique for physiological examination of canine skeletal muscle in vivo.

A technique is described for studying the physiological function of canine skeletal muscle in vivo. The contractile properties of the tarsal flexor muscles were examined in three beagle dogs under general anaesthesia. The force responses to electrical stimulation of the common peroneal nerve were measured at various frequencies to determine the frequency:force relationship for this muscle group. Fatigue characteristics were also examined during intermittent stimulated activity delivered in a set pattern of frequencies. The results provide quantitative characterisation of muscle function which is repeatable. The technique described could be applied to other animals and is a potentially powerful tool for evaluating the effects of drugs on muscle performance.

Animals↗
PubMed1988

Distribution of fibre types and fibre sizes in the tibialis cranialis muscle of beagle dogs.

The percentages of Type I muscle fibres were measured systematically in ATPase-stained, transverse cryostat sections of whole tibialis cranialis muscles from 8 young, adult beagles. The distance of the section from the origin of the muscle does not significantly affect the mean percentage. There are no identifiable differences in mean percentages between right and left muscles. Differences in mean percentages between individuals are significant when sexes are combined (P less than 0.01) and within sexes (males: P less than 0.01; females: P less than 0.05). Within sections, the percentage tends to be lowest at the superficial (craniolateral) border and to vary less from site to site deeper within the muscle. Fibre cross sectional areas were measured systematically in the same sections of the right muscle from 3 males and 3 females. Mean areas for each section were greater for Type II than for Type I fibres. Mean areas for each fibre-type varied moderately and non-systematically between the sample sites within sections. A needle biopsy taken from deep within this muscle should provide a more consistent and reliable estimate of fibre-type proportion in the whole muscle than a superficial specimen. Proportions are not affected by the distance of the sample site from the muscle origin, and left or right muscles are suitable for sequential samples.

Animals↗
PubMed1987

Myopathy with core-like structures in a dog.

Core-like structures were seen histologically in many of the fibres of the triceps and biceps femoris muscles of an 18-months-old male Great Dane with muscle weakness and moderate proximal muscular atrophy. The structures were lightly staining and lacked cross-striations. Some contained vacuoles and nuclei. Scattered necrotic and presumably regenerating fibres were also present. Ultrastructurally, the cores contained disarrayed filament bundles attached to thickened Z-lines which were compatible with the rods of rod myopathies. The condition was not fully characterized, but has certain similarities to a group of rare human congenital muscular disorders which includes central core disease.

Actin Cytoskeleton↗
PubMed1987

Acute Lantana camara toxicity in cattle.

An outbreak of acute Lantana camara poisoning in cattle is described in which 10 out of 91 animals died. The affected cattle became icteric and voided soft, black faeces. Necropsies were performed on three steers, and the macro- and microscopical changes in their livers and kidneys were compatible with those of L. camara poisoning. Changes were similar in two steers that developed typical signs after being dosed with fresh L. camara collected in the toxic camp. Clinical pathological changes in experimental animals included elevated serum urea and creatinine concentrations.

Animals↗
PubMed1985

An outbreak of cerebrocortical necrosis (polioencephalomalacia) in goats.

In a herd of 30 Boer goats, 3 young goats developed nervous signs including apparent blindness. In 2 of the goats a diagnosis of cerebrocortical necrosis was confirmed by the demonstration of lesions typical of the disease on histopathological examination. Lead concentrations in the renal cortex were well below the level regarded as indicative of lead poisoning in ruminants. The reason for the outbreak was not clear, but the feeding of concentrate and poor quality roughage may have been contributory factors. Cerebrocortical necrosis appears to be unusual in goats, compared to cattle and sheep, but it should be entertained in the differential diagnosis of caprine nervous diseases.

Animals↗
PubMed1985

A haemorrhagic syndrome in recently weaned pigs ascribed to hypovitaminosis K.

An outbreak of a haemorrhagic syndrome involved recently weaned, mixed-breed pigs in a large piggery. The pigs were fed a pelleted complete ration containing antibacterial drugs. Affected pigs failed to grow, became pale and developed large, subcutaneous haematomas. Some pigs became lame and one had epistaxis. The monthly mortality rate in the weaner house, which was previously less than 2%, exceeded 6% during the outbreak. Coagulation time, activated partial thromboplastin time and prothrombin time were prolonged in blood from some of the pigs. The outbreak resolved promptly after supplementation of the diet with vitamin K3.

Animals↗
PubMed1985

A suspected lipofuscin storage disease of sheep associated with ingestion of the plant, Trachyandra divaricata (Jacq.) Kunth.

Paresis afflicted 85 out of a flock of 770 young Merino ewes kept on old wheat lands in the western Cape during a period of drought. Many of the paretic ewes died. The vegetation was sparse and was dominated by Trachyandra divaricata. At necropsy, yellowish-brown discoloration of the grey matter throughout the brain and spinal cord and mild brown discoloration of the liver, renal cortex and lymph nodes were consistently seen. Light microscopical examination revealed abundant, yellowish-brown pigment granules in the cytoplasm of most of the larger neurons. Similar pigment also occurred in some non-nervous tissues. Shrinkage and loss of a few randomly scattered axons were observed in the white matter of the spinal cord in 2 sheep. Histochemical and ultrastructural features of the pigment were consistent with those of lipofuscin. T. divaricata failed to reproduce the condition when dosed to a sheep, but the paresis and pigmentation shown to be caused by the closely related plant, T. laxa, are strikingly similar. Trachyandra poisoning appears to be the first documented example in farm animals of an acquired lipofuscin storage disease involving nervous and non-nervous tissues for which a specific plant has been causally implicated.

Animals↗
PubMed1984

Myocardial pathology of domestic ruminants in southern Africa.

Myocardial pathology of ruminants in southern Africa, including lesions associated with toxic plants, other toxic agents, infectious agents and nutritional deficiency, is discussed with regard to recognition and to aetiological diagnosis. Findings are included which have not been published elsewhere. The importance and difficulties in recognition of myocardial lesions at an early stage are emphasized. Further research into the pathology of cardiac failure caused by toxic plants is clearly needed.

Animals↗
PubMed1984

Measurements of mass, length and valve diameters from normal formalin-fixed ovine hearts.

Hearts from 60 Merino sheep of known age, sex and live mass and with no known history of disease were collected and fixed in buffered 10% formalin. Systematic light microscopical examination did not indicate any abnormality in hearts of any of the sheep. The mass of various parts of the hearts, the length of the hearts and the diameters of the heart valves were measured to establish a basis for quantitative assessment of possible pathological changes associated with the ingestion of cardiotoxic plants. The mass measurements and, to a lesser degree, the lengths of the hearts varied considerably, but the ratio of the mass of the left ventricle plus ventricular septum divided by the right ventricular free wall mass was remarkably stable, and is promising as an indicator of right ventricular hypertrophy.

Age Factors↗