Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

Seasonal variation in alkalinity in pans in Central Africa

Summary The alkalinity and pH of lakes (pans) in Central Africa on Kalahari Sand, varies with the physical nature of the pan. Natural pans show a range of pH from 6.1 to 8.7, and of total alkalinity from 10 mg/l to 580 mg/l. This range is attributable to local soil differences. Some pans have been deepened or have had water pumped into them to provide drinking points for game animals. The alkalinity of pumped pans is determined by the alkalinity of the water pumped into the pan and the time during which this has taken place. All these types of pan show cyclic seasonal variation in alkalinity and pH. The range in these factors is similar from one year to the next. Animals living either in permanent (deepened or pumped) pans or in temporary (natural) pans have to withstand a wide range of alkalinity fluctuation in the course of the year.     

Asthma in children--problems in diagnosis

Bronchial asthma in children may be difficult to diagnose. Education of the parents regarding allergic conditions, specifically bronchial asthma, is exceedingly important in order to assure satisfactory treatment and clinical results. Chest symptoms of unexplained origin in early life should immediately arouse suspicion of allergic disease. Other causes of asthmatic symptoms must be borne in mind and excluded before a positive diagnosis of bronchial asthma is established. Of the many factors to be considered in investigating a child with asthma, a comprehensive history is most essential. The climate to which the patient is exposed and the psychic influences must be taken into account. Physical examination, x-ray films and laboratory procedures should be carefully executed. Skin testing, especially with food allergens, should not be relied upon to give all the information in allergic disease. Some form of diet trial, such as elimination diets, should be used if sensitivity to food is suspected.     

Oxygen tension in cerebral microvessels in acute anaemia in rats

Using polarographic oxygen microelectrodes, distribution of oxygen tension (pO2) in the rat cerebral arterioles (with a lumen diameter of 8-80 microm) and venules (with a lumen diameter of 8-120 microm) has been studied in acute reduction of haemoglobin concentration in the blood. Isovolumic haemodilution with 5 % albumin solution has been performed stepwise from 14 g/dl (control) to 10 g/dl (step 1), 7 g/dl (step 2) and to 4.6 g/dl (step 3). It was shown that step 1 of haemodilution led to no impairment of oxygen supply to the brain cortex. Step 2 resulted in moderate increase of pO2 in arterioles, whereas in venules oxygen tension fell down substantially (on the average, to 32 mm Hg). Step 3 resulted insignificant increase of pO2 in arterioles. A further fall of pO2 (to 27 mm Hg) in studied venules was recorded. The portion of venules with low pO2 grew to 31% (only 3 % in control). Microregions with a near-to-zero pO2 were recorded in some capillaries. This indicates presence of hypoxic zones in brain tissue. Hypoxic and anoxic microregions originate at this stage of anemia in locations with relatively low and/or impaired blood supply.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.     

Nuclear conditions in haploid Pelargonium in vivo and in vitro

In the shoot apices of the haploid Pelargonium cultivar Kleine Liebling, all mitoses are haploid (n = 9); however, ca. 20% of the interphase nuclei have DNA contents greater than 2C (up to 4C), indicating a tendency to chromosome endoreduplication in this material. — In internodes in vivo, the few mitoses present are haploid (quite probably, cambium cells); in addition to haploid interphases (1C to 2C DNA contents), endoreduplicated (endopolyploid) nuclei and nuclei in the course of endoreduplication occur with a frequency of ca. 40–50% (DNA contents up to 8C). — When internodes are cultured in vitro, differentiated cells are stimulated to divide, thus forming a population of diploid and tetraploid mitoses in addition to the preexistent meristem (haploid) cell population. In the process of time, diploid and tetraploid mitoses continue to be present in the callus, whilst haploid mitoses may decrease in number and eventually disappear. All mitoses analyzed had euploid chromosome numbers (9, 18 and 36) and their DNA contents were correspondingly 2C, 4C and 8C. Since no extensive chromosome counts were made, aneuploidy in the cultured material cannot be excluded; but, if occurring, it should be rather rare. — Under the experimental conditions used, prolonged culture in vitro leads to the production of nuclei with DNA contents (16C and 32C) greater than those occurring in vivo (8C), due to one and two additional DNA replications respectively beyond the limits attained in vivo. Even in these cultures, however, a population of the meristematic haploid cell line (DNA values 1C to 2C) is still present. — The present results are discussed in their relations with previous works on nuclear conditions in vivo and in vitro and on regeneration processes in cultured tissues in plants.     

Trends in dental caries in 12-year old children in Croatia

In this epidemiological study the caries status of 12-year-old children was compared over time for the years 1968, 1973, 1980, 1985, 1990, 1991 and 1999. The mean number of decayed, missing and filled teeth (DMFT) decreased from 7.0 in 1968 to 2.6 in 1991. From 1991 to 1999 and increase of DMFT 2.6 in 1991, to 3.5 in 1999) has been seen which is result of war in Croatia and reorganisation of health care delivery system. With the expected continuing decline in the prevalence and severity of oral diseases, one of the main problems in the future seems to be the transition of the oral health care delivery system from a disease-oriented system to a health-oriented one.     

Variation in colour-blindness in North-East India

Variation in colour-blindness in some of the populations of North-East India has been examined. The people of North-East India are found to be extremely variable with regard to this trait, the frequency ranging from 0 to 8.05%. An attempt has also been made to examine the present findings in light of the hypothesis put forward by Post (1962a,b) and others about selection relaxation.     

Stereoselective metabolism of pentoxifylline in vitro and in vivo in humans

Pentoxifylline increases erythrocyte flexibility, reduces blood viscosity, and inhibits platelet aggregation and is thus used in the treatment of peripheral vascular disease. It is transformed into at least seven phase I metabolites, of which two, M1 and M5, are active. The reduction of the keto group of pentoxifylline to a secondary alcohol in M1 takes place chiefly in erythrocytes, is rapidly reversible, and creates a chiral center. The aims of this study were: to develop HPLC methods to separate the enantiomers of M1, to investigate the kinetics of the reversible biotransformation of pentoxifylline to (R)- and (S)-M1 in hemolysed erythrocyte suspension, and to quantify the formation of the enantiomers of M1 (as well as M4 and M5) after intravenous and oral administration of pentoxifylline to human volunteers. (R)- and (S)-M1 could be separated preparatively on a cellobiohydrolase column, while determination in blood or plasma was by HPLC after chiral derivatization with diacetyl-L-tartaric acid anhydride. The metabolism of pentoxifylline to (R)-M1 in suspensions of hemolysed erythrocytes followed simple Michaelis-Menten kinetics (K(m) = 11 mM), while that to (S)-M1 was best described by a two-enzyme model (K(m) = 1.1 and 132 mM). Studies with inhibitors indicated that the enzymes were of the carbonyl reductase type. At a therapeutic blood concentration of pentoxifylline, the calculated rate of formation of (S)-M1 is 15 times higher than that of the (R)-enantiomer. Back-conversion of M1 to pentoxifylline was 3-4 times faster for the (S)- than for the (R)-enantiomer. In vivo, the R:S plasma concentration ratio of M1 ranged from 0.010-0.025 after intravenous infusion of 300 or 600 mg of pentoxifylline, and from 0.019-0.037 after oral administration of 600 mg. The biotransformation of pentoxifylline to M1 was thus highly stereoselective in favor of the (S)-enantiomer both in vitro and in vivo.     

Scrapie in Sheep in Sweden

In 1986 scrapie was diagnosed in 2 ewes of Swedish landrace (finn sheep) from a herd south of Stockholm (Carlsson et al 1986). As the diagnosis was based on clinical histo-ry and patho-anathomical changes only, inoculation tests in mice and goats were per-formed to try to verify the diagnosis.     

Chemokines in host-parasite interactions in leishmaniasis

Crucial to the defense against leishmaniasis is the ability of the host to mount a cell-mediated immune response capable of controlling and/or eliminating the parasite. Cell recruitment to the site of infection is essential to the development of the host cellular immune response. The process is controlled by chemokines, which are chemotactic cytokines produced by leukocytes and tissue cells.     

Pyruvate oxidation in vivo and in vitro in Dictyostelium discoideum

The in vivo rate of the reaction catalyzed by the pyruvate dehydrogenase complex in Dictyostelium discoideum was measured by exposing cells to [1-14C]alanine and relating the rate of 14CO2 evolution to the specific radioactivity of the isolated [1-14C]pyruvate. The rate obtained, 0.33 mumol/min per ml packed cell volume, was very similar to that determined by independent calculations based upon the rate of utilization of endogenous protein as an energy source in this system.     

Scurvy in capybaras bred in captivity in Argentine

In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity.     

Prospects for in vivo NMR methods in xenobiotic research in plants

The application of non-invasive nuclear magnetic resonance (NMR) methods in xenobiotic research is reviewed in relation to: (i) the characterisation of the effects of xenobiotics on the metabolism of plants and plant cell suspensions; (ii) the direct detection of xenobiotics and their degradation products in vivo; and (iii) the spatial localisation of xenobiotics and their derivatives at the subcellular and tissue levels. Novel information has been generated by in vivo NMR studies of both agrochemicals and heavy metals, but a lack of generality in the methods makes it difficult to extrapolate from one successful application to the next. In vivo NMR spectroscopy is shown to be informative when a xenobiotic perturbs metabolic pathways that are accessible to the technique, and it is useful for probing the partitioning of paramagnetic metal ions between the cytoplasm and the vacuole. The successful application of 19F NMR to the analysis of plant tissue extracts also suggests that in vivo 19F NMR spectroscopy may have a role in biotransformation studies of fluorinated xenobiotics. In contrast NMR imaging techniques have been little used for xenobiotic research in plants, and while the method has been shown to be capable of monitoring the uptake and translocation of paramagnetic ions in plants, the potential use of high resolution 1H and 19F NMR imaging for mapping agrochemicals in tissues is still in its infancy.     

Meiosis in Bovine Oocytes Matured in Vitro and in Vivo

Meiosis in bovine oocytes has; been studied after maturation in vitro or in vivo. Oocytes for in vitro maturation were collected from the ovaries of slaughtered cattle without regard to the phase of the estrous cycle while in vivo maturation was studied in oocytes from gonadotrophin-stimulated heifers at times varying between 6 and 36 h after the beginning of behavioural estrus. Oocytes from slaughtered cattle were classified according to their cumulus complex and ooplasm and were cultured for 6, 12, 18, 24, 36 or 48 h in modified Krebs-Ringer bicarbonate buffer before fixation) for cytogenetic analysis. Oocytes from stimulated heifers were aspirated from follicles or flushed from the oviducts, classified according to cumulus and ooplasm, and fixed within 6 h of collection. Nuclear maturation was more rapid in vitro than in vivo. The largest proportion of oocytes reached maturity (Mil) after 12 to 18 h in culture or 30 to 36 h after the onset of behavioural estrus. Oocytes devoid of cumulus cells or showing signs of vacuolation or degeneration had virtually no capacity for nuclear maturation.     

Biomarkers discovery in cancer--up-dates in methodology

Biomarkers are biomolecules that can indicate normal/pathological processes, or physiological responses to therapy. Due to the serum abundance in proteins, such as albumin and lypo/glycoproteins, biomarkers are difficult to assess. Serum biomarkers identification can contribute to personalized medicine and improve cancer diagnostic and prognostic. The paper summarizes some of the proteomics techniques and the workflow used for protein signatures identification associated to cancer development. Thus, biomarkers validated for prostatic, breast, cervical or lung cancers are presented as examples for clinical application of serum markers. In spite of the continuous research efforts, there are only few validated biomarkers that have proved a good predictive power in cancer. Modern technology and the combination of various techniques used for proteins quantification represent important means for the identification and validation of new biomarkers.     

Directional cues in tactile stimuli involved in agonistic encounters in cockroaches

ABSTRACT. The ability of male cockroaches, Nauphoeta cinerea and Periplaneta americana , to respond directionally to tactile agonistic acts was tested using stimulation by artificially manipulated appendages. Responses by Nauphoeta included turning towards the stimulus at preferred angles of c. 40°, 90° and 180°, apparently relying on internally-stored directional sensory information. This turning responsiveness depended in part on the social status of the receiving individual, since subordinate individuals often retreated or failed to respond. Periplaneta males reacted to tactile stimuli by quick movements away from the stimulus or by kicking towards it. The leg used in kicking was that nearest to the part of the body which was stimulated.     

Salmonella in Sheep in Iceland

In 1995 several outbreaks of food poisoning in humans occurred in Iceland, that were traced to salmonella contamination of singed sheep heads. This prompted us to study the prevalence of salmonella infection in sheep and to trace where and how infection might have occurred. Faecal, intestinal contents and tonsillar samples were collected in the spring and autumn from sheep on 50 farms in the southwestern part of the country, where salmonellosis had been detected and from 5 farms in the northwestern part of the country. All faecal samples from the southwest were negative, whereas samples from 3 farms obtained in the autumn in the northwest were positive. Tonsillae taken in the autumn were positive in sheep from 3 farms in the southwest and 2 in the northwest. Our results show that salmonella infection is rare in Icelandic sheep but healthy carriers may harbour the bacteria in tonsillae. Salmonella was not detected in drainage from slaughterhouses nor in singed sheep heads.