Isolation and characterization of butanol-resistant mutants of Clostridium acetobutylicum.

In a wild-type strain of Clostridium acetobutylicum isolated from soil, solvent production appeared limited by butanol toxicity. Butanol-resistant mutants have been obtained which produced significantly higher solvent concentrations (about 30%) than the wild-type strain. Some other physiological differences were observed between a selected resistant mutant and the wild-type strain at the level of solvent resistance and sporulation.     

Cytochemical study of macrophage lysosomal inorganic trimetaphosphatase and acid phosphatase

Cytochemical investigations have associated acid inorganic trimetaphosphatase (TMPase) activity with the lysosomes of certain cell types. We have used the modified staining technique of Berg to show that this enzyme activity is present in normal mononuclear phagocytes and macrophage cell lines. We have found this enzyme activity to be present in murine RAW264 macrophages, in human U937 macrophages, in normal human blood monocytes, and in guinea pig peritoneal macrophages. All of the RAW264 and U937 macrophages showed intense TMPase activity. Many of the human monocytes and most of the guinea pig macrophages were labeled by this method. The reaction product was associated with the lysosomes of these cell types. The lysosomal staining-pattern was similar to that of acid phosphatase. Differences with regard to Golgi staining were noted. This indicates that TMPase is a lysosomal enzyme of mammalian macrophages. The distinction between TMPase and acid phosphatase activity has been demonstrated by measuring the pH optimum of each enzyme. Using substrates identical to those of the ultrastructural cytochemistry, we show that the pH optimum of TMPase is 4.0 and that of acid phosphatase is 5.0. The enzymatic activities are therefore ultrastructurally and biochemically distinct. Following phagocytosis of latex, yeast (Saccharomyces cerevisiae), or Corynebacterium parvum, TMPase has been found to be associated with phagosomes. This enzyme may take part in the degradation of phagocytosed materials, particularly microorganisms which contain inorganic polyphosphates and metaphosphates.     

Zymomonas mobilis mutants blocked in fructose utilization

Summary A mutant ofZymomonas mobilis deficient in the utilization of fructose for growth and ethanol formation was shown to lack fructokinase activity. When grown in media which contained glucose+fructose or sucrose, both the mutant and wild type produced sorbitol in amounts up to 60 g·l^-1, depending on the initial concentrations of sugars. Sorbitol formation was accompanied by an accumulation of acetaldehyde, gluconate, and acetoin. A ferricyanide-dependent sorbitol dehydrogenase could be localized in the cell membrane; it thus resembles the sorbitol dehydrogenase ofGluconobacter suboxydans. Neither a NAD(P)H dependent reduction of fructose nor a NAD(P) dependent dehydrogenation of sorbitol could be detected in cell-free extracts. The use of fructose-negative mutants ofZ. mobilis for the enrichment of fructose in glucose+fructose mixtures is discussed.     

Angiotensin biosynthesis and concentrations in brain of normotensive and hypertensive rats

We report here on the extraction and characterization of angiotensin I (ANG I) and angiotensin II (ANG II) from the brain of rats. High pressure liquid chromatography (HPLC) with different mobile phases combined with specific radioimmunoassays (RIA) proved to be a powerful tool for peptide characterization in biological samples; (Ile5)-ANG I, (Ile5)-ANG II and (Ile5)-ANG III could clearly be identified in cerebrospinal fluid (CSF), incubated in vivo and in vitro with renin, in total brain extracts, as well as in hypothalamus (HT), medulla oblongata (MO), cerebellum (CER) and cortex (CO). Angiotensin cleaved from CSF angiotensinogen and angiotensin extracted from brain showed retention times identical to those of plasma angiotensin and synthetic standard peptides, indicating that their amino acid sequence is probably identical. ANG I and ANG II were highest in the HT and lowest in the CO. Following bilateral nephrectomy (NX) both ANG I and ANG II persisted at control levels. Young 10 week old spontaneously hypertensive rats (SHRSP) showed significantly lower ANG I and ANG II concentrations in the HT compared with Wistar Kyoto rats (WKY). Intracerebroventricular (i.c.v.) administration of the converting enzyme inhibitor captopril caused a significant increase in ANG 1 in nephrectomized SHRSP but not in WKY. These differences were not found in 40 week old SHRSP. The data show that ANG I and ANG II are synthetized in the brain of rats. The lower concentrations and the enhanced accumulation of ANG I after converting enzyme blockade in nephrectomized young SHRSP indicate an increased turnover of angiotensin in hypertensive rats.     

Investigation of cellobiohydrolase from trichoderma reesei by small angle X-ray scattering

Summary Trichoderma reesei was grown on sulfite pulp and the major cellobiohydrolase of the culture filtrate was purified to homogeneity. The distance distribution function p(r) measured by the small angle X-ray scattering technique indicates that the enzyme molecule has a rather unusual tadpole like shape with an isotropic head and a long tail. The maximum length is 18 nm and the largest diameter is 4.4 nm.     

Control of echolocation pulses by neurons of the nucleus ambiguus in the rufous horseshoe bat,Rhinolophus rouxi

Horseradish peroxidase was applied by inotophoretic injections to physiologically identified regions of the laryngeal motor nucleus, the nucleus ambiguus in the CF/FM bat Rhinolophus rouxi. The connections of the nucleus ambiguus were analysed with regards to their possible functional significance in the vocal control system, in the respiration control system, and in mediating information from the central auditory system. The nucleus ambiguus is reciprocally interconnected with nuclei involved in the generation of the vocal motor pattern, i.e., the homonomous contralateral nucleus and the area of the lateral reticular formation. Similarly, reciprocal connections are found with the nuclei controlling the rhythm of respiration, i.e., medial parts of the medulla oblongata and the parabrachial nuclei. Afferents to the nucleus ambiguus derive from nuclei of the 'descending vocalization system' (periaqueductal gray and cuneiform nuclei) and from motor control centers (red nucleus and frontal cortex). Afferents to the nucleus ambiguus, possibly mediating auditory influence to the motor control of vocalization, come from the superior colliculus and from the pontine nuclei. The efferents from the pontine nuclei are restricted to rostral parts of the nucleus ambiguus, which hosts the motoneurons of the cricothyroid muscle controlling the call frequency.     

Interferon induction by platinum(II)-poly(I).poly(C) complexes

The effect of the interaction between poly(I).poly(C) and cis-dichloro-diammineplatinum(II) (cis-Pt), its trans analogue and chloro-diethylene-triamminoplatinum(II) (dien-Pt) on interferon induction activity was investigated. The covalent monodentate fixation of the three compounds on N7 of inosine has different effects on the structure and thermostability of poly(I). poly(C) which is well reflected by the interferon induction activity of the samples. Thus, the sandwich stabilization by dien-Pt at low binding ratios is manifested by an increased interferon induction and a high resistance towards RNAase degradation. The destabilization of the duplex by cis-Pt decreases interferon induction, accompanied by an increase in RNAase sensitivity of the complexes. In the case of trans-Pt the duplex structure is little perturbed and interferon induction is essentially maintained.     

Zur Wirkung des Lichtes auf die circadiane Periodik des Stoffwechsels und der Aktivität beim Buchfinken (Fringilla coelebs L.)

Zusammenfassung Bei 8 Buchfinken (4, 4) wurden Sauerstoffverbrauch und Hüpfaktivitätsowohl unter Bedingungen eines künstlichen Licht-Dunkel-Wechsels (LD 1212 Std; ohne Dämmerung) als auch im Dauerlicht (LL) bei konstanter Umgebungstemperatur (18–20° C) kontinuierlich gemessen. Im 1. Teil des Versuchs wurde die Beleuchtungsstärke während der Dunkelzeit (D) zwischen 0,01 und 1,0 Lux bei gleichbleibender Beleuchtungsstärke während der Lichtzeit (10 Lux) geändert. Im 2. Teil des Versuchs wurden die Vögel im Dauerlicht von 1 und 10 Lux untersucht. Vier Vögel wurden zusätzlich bei 100 Lux gemessen.Das mittlere Niveau des Sauerstoffverbrauchs (pro Std) und die mittlere Aktivitätsmenge (pro Stunde) während einer Periode sind bei der Mehrzahl der untersuchten Vögel linear und positiv miteinander korreliert (Abb. 2 und 3; Tabelle 1).Änderungen des mittleren Aktivitätsumsatzes (Differenz von mittlerem Niveau des Gesamtumsatzes und des Ruheumsatzes) sind ebenfalls positiv mit Änderungen der Aktivitätsmenge korreliert. Diese Aussage beruht auf der Voraussetzung, daß sich die circadiane Periodik des Gesamtstoffwechsels aus mindestens zwei Periodizitäten zusammensetzt: der Periodik des Grundstoffwechsels (vegetative Periodik) und der Periodik von Ruhe und Aktivität (Aktivitätsperiodik).Bei Änderungen des Zeitgebers (Erniedrigung der Zeitgeberamplitude und gleichzeitige Erhöhung der mittleren Beleuchtungsstärke) werden folgende Parameter signifikant verändert: die positive Phasenwinkel-Differenz zwischen Stoffwechselperiodik und Zeitgeber wird vergrößert, die Dauer der Aktivität (Aktivitätsumsatz ₃ ) wird verlängert, die Schwingungsbreite der Stoffwechselperiodik wird erniedrigt (Abb. 4; Tabelle 2).Unter Dauerlicht-Bedingungen werden durch eine Erhöhung der Beleuchtungsstärke von 1 auf 10 Lux folgende circadianen Parameter erhöht: die Frequenz, das mittlere Niveau der Stoffwechselperiodik (Gleichwert), der Ruheumsatz, das -Verhältnis (Verhältnis Aktivitätszeit/Ruhezeit) und die Aktivitätsmenge pro Zeiteinheit. Die Schwingungsbreite der Stoffwechselperiodik bleibt unverändert (Abb. 5). Außerdem besteht eine regelhafte Beziehung zwischen dem Formfaktor der Stoffwechselschwingung und der Beleuchtungsstärke, beziehungsweise der Spontanfrequenz. Erhöhung der Beleuchtungsstärke von 10 auf 100 Lux hat unterschiedliche Wirkungen auf die circadianen Parameter der 4 gemessenen Vögel.Die unter Dauerlicht-Bedingungen gewonnenen Ergebnisse folgen der modifizierten circadianen Regel, die eine positive Korrelation zwischen Frequenz und Gleichwert der autonomen circadianen Schwingung fordert (Abb. 7).Die unter Zeitgeber-Bedingungen (Licht-Dunkel-Wechsel) gewonnenen Ergebnisse werden auf der Grundlage einer vorwiegenden Beeinflussung des circadianen Schwingers durch die Amplitude der Zeitgeber-Periodik (Zeitgeberstärke) gedeutet. Änderungen der Beleuchtungsstärke während der Dunkelzeit (im Bereich der gebotenen Lichtintensitäten) haben keinen regelhaften Einfluß auf die Stoffwechselparameter der Periodik.

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On the effect of light upon the crcadian rhythms of metabolism and activity in the chaffinch (Fringilla coelebs L.)

Summary Oxygen consumption and hopping activity of 8 chaffinches (4 males, 4 females) were measured simultaneously (I) in an artificial 1212 hr light-dark-cycle (LD; without twilight) and (II) in continuous illumination, both at constant temperature (18–20°C). During the first part of the experiment illumination during darktime (D) was varied in both directions between 0.01 and 1.0 lux. Light intensity in L (lighttime) remained constant (10 lux). In the second part of the experiment the birds were exposed to a continuous illumination of 1 and 10 lux. Four birds were also measured at 100 lux.For the majority of the birds, the mean level of oxygen consumption per hour and the average amount of activity (contacts per hour) during one circadian period are positively and linearly correlated to each other (Fig. 2 and 3; Table 1).Changes in the level of the activity metabolism — i.e. the difference between mean level of total metabolism and resting metabolism — are also positively correlated to changes in the amount of activity. This statement is based on the assumption that there exist at least 2 rhythms as basic components of the circadian rhythm of total metabolism: the rhythm of basal metabolic rate (BMR) or vegetative rhythm and the rhythm of activity and rest (activity rhythm).By variation of the Zeitgeber — simultaneous decrease in amplitude and increase in mean level of light intensity — the following parameters are significantly changed: the phase-angle difference between the circadian rhythm of metabolism and the Zeitgeber is increased (positive phase-angle differences are changed in positive direction), the activity time is lengthened, and the amplitude of the rhythm (range of oscillation) is diminished (Fig. 4; Table 2).With an increase in light intensity from 1 to 10 lux (continuous illumination), the following parameters are increased: the frequency of the rhythm, the mean level of metabolism per period, the resting metabolic rate, the -ratio (i.e. ratio activity time/rest time), and the total amount of activity per unit time. The range of oscillation remains unchanged (Fig. 5). There is also a regular change of the form-factor characterizing the shape of the metabolic oscillation, which depends upon light intensity as well as upon frequency (Fig. 8). Increasing light intensity from 10 to 100 lux affected differently the circadian parameters of the four birds tested.The results obtained in continuous illumination follow the modified circadian rule which postulates the positive correlation between frequency and mean level of the autonomous circadian oscillation (Fig. 7).The results obtained under the influence of a Zeitgeber (light-dark-cycle) are interpreted on the basis of the primary influence of the Zeitgeber amplitude (strength of the Zeitgeber) upon the circadian oscillation. The given changes in light intensity during darktime have no significant effect on the metabolic parameters of the rhythm.