Understanding the mechanism of action of Cytochrome c Oxidase in normal and disease states: shark heart enzyme, a potential model

Cytochrome c oxidase, the final member of the electron transport chain, is crucial to respiration and also contributes to the synthesis of cellular ATP. The total absence of this enzyme is incompatible with life and its deficiency or malfunction leads to a number of serious disease states. Understanding the mechanism of action of this enzyme, which is an important prerequisite to unravelling its role in the pathogenesis of disease states, is hampered by the lack of suitable enzyme models. The bovine enzyme, which are structually simple, appear to follow a different mechanism of action. The hammer head shark is a seasonal resident of the warm waters of the Caribbean Sea. The work presented here indicates that, like the bovine enzyme, the enzyme of the heart of this shark (i) possesses thirteen subunits and two substrate binding sites and (ii) exhibits biphasic kinetics. The work also confirms that, unlike the bovine enzyme which is dimeric, the shark enzyme functions as a monomer. Given this latter simplifying feature, in conjunction with its kinetic and structural similarities to the more complex mammalian varieties, we propose that shark heart cytochrome c oxidase replace the bovine and bacterial forms as the enzyme of choice for model studies.(Au)

The feasibility of a porcine model of acute coronary occlusion and reperfusion using off-pump coronary artery bypass grafting

We developed an open-chest porcine model of acute coronary occlusion and surgical reperfusion, and attempted to prevent intra-operative ischaemic ventricular fibrillation (VF) by a Retrograde Intracoronary Glyceryl trinitrate (RIG) infusion into the occluded vessel. Five Yorkshire pigs (weight 50ñ 1.1kg), randomized into 3 groups, underwent median sternotomy under general anaesthesia. One pig (Group 1, control) underwent sternotomy and pericardiotomy only. Four pigs underwent acute left anterior descending (LAD) coronary occlusion. Two pigs were not reperfused (Group 2). Two pigs underwent surgical reperfusion (Group 3) via left internal mammary artery (LIMA) grafting to the LAD using the Off-Pump Coronary Artery Bypass (OPCAB) technique. Ischaemic injury was assessed using 7-lead electrocardiography (EGG) and transthoracic/epimyocardial echocardiography (ECHO). Group 1: transient intraoperative hypotension and VF occurred. Successful resuscitation and 10-week survival (until sacrifice) with normal left ventricular (LV) function was achieved. Group 2: there were ECG and ECHO evidence of acute LV ischaemic dysfunction in both pigs. The surviving pig had persistent anterior hypokinesis at 8« months. The other died intra-operatively following progressive ischaemic LV dysfunction despite resuscitative attempts. Group 3: the surving pig had normal LV function at 8 months. Initial anterior LV akinesis normalized within 7 days. The other developed post-occlusion haemodynamic instability and died intra-operatively despite reperfusion. In this porcine model, acute LAD artery occlusion modified by the novel RIG infusion technique, followed by surgical reperfusion (OPCAB) is feasible. This model would facilitate further development of OPCAB surgical expertise and understanding of the pathophysiology of ischaemia-reperfusion injury.(Au)

An experimental mouse model to study the pathogenicity of Prevotella bivia and investigations of possible virulence

Induction of subcutaneous abscesses in mice was used to study the pathogenicity of Prevotella bivia both in mono-infection and in mixed cultures with Escherichia coli and Peptostreptococcus spp. Virulence factors such as coaggregation and aggregate formation of cells, haemagglutination activity and tolerance to serum bactericidal activity were investigated for their possible role in P bivia pathogenicity. Monocultures of P bivia, E coli and Peptostreptococcus spp did not induce subcutaneous abscess at concentrations as high as 10 9 colony forming units/millilitre (cfu/ml). Only E coli persisted at the infection site for up to 7 days post infection but with a marked decline in cell count (8.0x10 squaredcfu/ml). The anaerobic organisms did not persist at the infection sites beyond the fifth day. In contrast, mixed cultures of P bivia and E coli or all three organisms potentiated for infective abscess two weeks after infection. Viable cells were recovered from abscesses in greater numbers as the infection progressed. Prevotella bivia was the predominant organism in chronic abscesses while E coli predominated in abscesses in the acute stage of the infection. Prevotella bivia lacked haemagglutination activity against human and sheep erythrocytes and showed marked susceptibility to 50 percent human serum. These may limit its haematogenous spread. Its ability to form aggregates in molar salt solutions and coaggregate with facultative organisms may account for its persistence in pathological sites.(Au)

Neuroprotection by caffeine and pentoxifylline during experimental cerebral ischaemia

Cerebral ischaemia was induced in anaesthetized rats by occlusion of the ipsilateral common carotid and middle cerebral arteries. The response to ischaemia was assessed by the reduction of the amplitude of recorded somatosensory evoked potentials (SSEPs), and the rate of recovery of the SSEPs during reperfusion. Caffeine and pentoxifylline when applied at 70 mM to the cortex for 60 min prior to induction of ischaemia significantly reduced the ischaemia induced attenuation of the SSEPs and hastened recovery to control levels. In contrast, application of normal saline or of the drugs for 15 min did not reduce the effect of ischaemia on the SSEPs. These results suggest that caffeine and pentoxifylline have potential roles in the management of patients with cerebral ischaemia.(AU)

Atopic dermatitis and HTLV-1 associated-myelopathy: associated or coincidental disorders?

Reports from Jamaica have indicated that some patients with infective dermatitis or atopic dermatitis (AD) are seropositive for antibodies to human T-lymphotropic virus type 1 (HTLV-1). We describe a 32-year-old Israeli woman with long-term AD and paresthesia in the distal parts of the extremities. Neurological examination revealed a positive Babinski's sign. HLA typing demonstrated that this patient has the common HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and infective dermatitis haplotype for DRBI* DQBI*. The presence of HTLV-1 was demonstrated with polymerase chain reaction; HTLV-1-antibodies were detected by the Western blot method and by inoculation of the patient's peripheral blood mononuclear cells into F344 rats. This study confirms the presence of HTLV-1 antibodies and proviral genome in a patient with AD which later evolved into HAM/TSP. We cannot yet conclude whether these two diseases are associated or coincidental disorders.(Au)

Plasmodium falciparum infection of splenectomized and intact Guyanan Saimiri monkeys

Spleen-intact and splenectomized Saimiri monkeys of Guyanan origin were examined for their potential suitability for Plasmodium falciparum protection studies. The animal could be readily infected with adapted strains of P. falciparium (Indochina 1/CDC and Uganda Palo Alto FUP strains), but spontaneously recovered without drug treatment and without development of severe clinical disease. In intact animals, peak parasitemia prior to recovery generally ranged from 0.1 percent to 10 percent, whereas in splenectomized animals the peak parasitemia was generally higher so that some animals were given drug treatment to assist in recovery from infection. In reinfection studies, previously infected spleen-intact monkeys demonstrated sterile immunity to the homologous parasite strain but not to a heterologous strain. However, in monkeys infected with the heterologous strain, the peak parasitemia was less than in the first infection and of shorter duration. Splenectomized animals did not demonstrate sterile immunity although the peak parasitemia achieved was less than in the previous infection of each of these monkeys. While the lack of major clinical disease indicated that these monkeys did not provide a good animal model for human malaria, the development of protective immunity was consistent with a useful role in evaluating candidate vaccine antigens (AU)

Human T cell lymphotropic virus: necessity for and feasibility of a vaccine

Human T cell lymphotropic virus types I and II (HTLV-I/II) are endemic in certain areas of the world. The cause two life-threatening diseases, adult T cell leukaemia/lymphoma and tropical spastic paraparesis. A vaccine is needed because in developing countries there are no other feasible preventive interventions against these diseases and in Western countries intravenous drug users at high risk for HTLV-I and HTLV-II infections and the health workers in contact with such populations must be protected. We have developed a rat model in which we observed variations of susceptibility to viral infection between inbred strains, the most susceptible being Fischer F344, and the possibility of viral latency in the nervous system. We have prepared a recombinant adenovirus vector that expresses the HTLV-I envelope glycoprotein env in HeLa cells. A target human population in French Guyana, in which the prevalence rate reaches 5.6 percent in one ethnic group (Bonis), has been identified for possible intervention (AU)

Clinical and biochemical features of malnutrition related diabetes mellitus

The aims of this study were to characterize clinically and biochemically malnutrition related diabetes mellitus (MRDM) as it presents in Jamaica (J-type or phasic insulin dependent diabetes mellitus) and to examine the food toxin (linamarin)/malnutrition hypothesis as a possible aetiological factor in the onset of MRDM. The study was broadly divided into two major areas: (i) Clinical or patient study and (ii) Animal model study. The patient population consisted of thirty-four, (23 females, 17 males) who attend the Diabetic Outpatient Clinic of the University Hospital of the West Indies. This group consisted of 14 phasic insulin dependent diabetes mellitus (PIDDM) (8 females, 6 males) 10 non-insulin dependent diabetes mellitus (IDDM), (6 females, 4 males). Ten (10) normal (4 females, 6 males) subjects also participated. The diabetic and normal subjects were matched for age, body mass index and duration of diabetes (among diabetics). Comparitive studies were performed on insulin receptor binding, hormonal profile, glucose tolerance, blood status, renal, hepatic and pancreatic function. The results show a significantly decreased white and red blood cell binding to insulin (P<0.05), extensive kidney damage (P<0.05) and increased pancreatic echogenicity in PIDDM. These findings support a separate identity of the latter syndrome from Types 1 and 11 diabetes mellitus. In the animal model study, the dogs (male and female) were maintained on a diet of cornmeal cooked with chicken, fortified with Purina laboratory chow and water ad libitum. Malnutrition was induced by restricting the diet to a very small quantity of cornmeal only for a period of 7-10 days. The recovered dogs were re-fed the normal diet with added milk and multi-vitamin supplement. Linamarin dosage of 20 mg per kg body weight induced abnormally high glucose levels, and in two cases the hyperglycaemia was sustained for several days. There was an associated decrease in binding of insulin to erythrocytes and mononuclear leucocytes, as well as prevailng hypoinsulinaemia in the limarin-induced hyperglycaemic conditions. This animal study presents a possible aetiological model for malnutrition related diabetes mellitus (Summary)

Studies on hypoglycin toxicity in rats. I. Changes in hepatic ultrastructure

Hypoglycin is a toxic amino acid found in the unripe ackee fruit. The ackee is a popular item of diet in Jamaica and has been proposed as a cause of the so-called vomiting sickness. Hypoglycin is thought to act by inhibiting the oxidation of long-chain fatty acids, uncoupling oxidative phosphorylation, and interfering with gluconeogenesis. Hypoglycin was given intraperitoneally to rats in a dose of 10 mg/100 g, and samples of liver taken at hourly intervals up to 5 hr were studied with the electron microscope and compared with controls. The major ultrastructural findings in the hypoglycin-treated rats were progressive mitochondrial swelling with loss of granules and pallor of the matrix, followed by incorporation into autophagic vacuoles. These findings correlate well with the reported biochemical mechanisms.(AU)