The beta1 integrin subunit is not a specific component of the costamere domain in human myocardial cells

Studies on altered integrin receptor expression during cardiac hypertrophy and heart failure requires accurate knowledge of the distributional pattern of integrins in myocardial cells. At present the general consensus is that in cardiac muscle the ₁ integrin receptor is mainly localized to the same sarcolemmal domain as vinculin at Z-band levels (costamere). Since most previous studies have been focusing on myocardial integrin distribution in lower mammals, the myocardial localization of the ₁ integrin subunit was investigated in biopsies collected from the auricle of patients undergoing a coronary bypass operation. Non-invasive serial optical sectioning was carried out by immuno-laser scanning confocal microscopy. Double-labelling for vinculin/-actinin, and the cytoplasmic domain for the ₁ integrin subunit, showed that ₁ integrin is deposited throughout both the vinculin/-actinin domains and the non-vinculin/-actinin domains. These results were supported by a semi-quantitative analysis in extended focus images of the latter preparations. Higher magnification views at the electron microscopical levels of the large, extracellular domain of the ₁ integrin subunit disclosed a pronounced labelling in the form of a dense, irregular punctuate pattern that was distributed at Z-disc domains as well as along the entire sarcolemmal area between Z-discs. Our findings show that in human, myocardial cells, the ₁ integrin receptor does not only localize to the surface membrane at the Z-disc level (costamere in cardiac muscle), but has a widespread distribution along the sarcolemma.     

A relationship between efficiency of isomaltosaccharide hydrolysis and thermostability of six Bacillus oligo-1,6-glucosidases

Summary A p-nitrophenyl--d-glucopyranosidase from Bacillus thermoamyloliquefaciens KP 1171 capable of growing at 30°–66°C was assigned to an oligo-1,6-glucosidase (dextrin 6--d-glucanohydrolase, EC 3.2.1.10). The enzyme was compared with its homologous counterparts from B. cereus NY-14, B. cereus ATCC 7064 (each mesophile), B. coagulans ATCC 7050 (facultative thermophile), B. thermoglucosidasius KP 1006 (DSM 2542, obligate thermophile) and B. flavocaldarius KP 1228 (extreme thermophile) in thermostability and kinetic parameters at suboptimal temperatures for isomaltosaccharides (2–6 glucose units). This analysis showed that the efficiency of each isomaltosaccharide hydrolysis changes in a convex manner with increasing thermostability on the transition, NY-14 ATCC 7064 ATCC 7050 KP 1071 KP 1006 KP 1228 enzymes, with a maximum at KP 1071 or ATCC 7050 enzyme.Presented at the Annual Meeting of the Agricultural Chemical Society of Japan, Kyoto, April 1, 1986     

Bifurcations of nonlinear reaction-diffusion systems in oblate spheroids

Spontaneous pattern formation may arise in biological systems as primary and secondary bifurcations to nonlinear parabolic partial differential equations describing chemical reaction-diffusion systems. Bipolarity in mitosis and cleavage planes in cytokinesis may be related to this formation of prepatterns. Three dimensional prepatterns are investigated, as they emerge in flattened spheres (i.e. oblate spheroids). Pattern sequences and selection rules are established numerically. The results confirm previously recorded results of the spherical and prolate regions, upon which a prepattern theory of mitosis and cytokinesis is based. Especially, the phenomenon of 90 degree axis tilting and the formation of a highly symmetrical saddle shaped pattern, crucial for the prepattern theory of mitosis and cytokinesis, is examined. Present results show, that these phenomena are stabilized in oblate spheroids. The bipolar mitosis prepattern is found as well, although the polar axis may appear with an angle toward the axis of the oblate spheroid. These results are thus further support for the prepattern theory of mitosis and cytokinesis.     

The entry process as the target for energy input in active transport of α-aminoisobutyric acid byMycobacterium phlei

Mycobacterium phlei was shown to accumulate -aminoisobutyric acid, establishing a concentration gradient of approximately 15,000-fold. The apparent affinity constant of the carrier for -aminoisobutyric acid was 1.8 µM. The system exhibited a broad specificity provided two structural requirements were satisfied: the presence of a free amino and carboxyl group on the alpha carbon and the absence of a net charge. The role of energy coupling on the accumulation of -aminoisobutyric acid was studied by two different kinds of experiments, the relative effects of the inhibitors on the rate of entry and the steady-state of accumulation, and a comparison of the efflux induced at the final steady state by the addition of (a) excess nonradioactive -aminoisobutyric acid, (b) energy inhibitors, or (c) both. The results are consistent with the hypothesis that accumulation of -aminoisobutyric acid is due to an increased rate of entry, the rate of exit not being affected by metabolic inhibitors.     

Oriented arrays of epicuticular wax plates inEucalyptus species

Summary Patterns of arrangement of epicuticular wax platelets are shown in a number of species ofEucalytpus. In some, wax platelets are oriented across the long axes of epidermal cells overlying veins and, in some species, over cells of adjacent stomatal complexes. Among other patterns, radial arrangements around stomata, arrangements in lines at right angles to underlying cell walls and concentric arrays are described. The possibility of the involvement of ectodesmata (ectocythodes) in the location of wax platelets is raised. The pattern of orientation of the platelets is ascribed to a prepattern, as yet unobserved, of anisotropic domains in the outer cuticle. It is suggested that this prepattern may consist of regions of molecular anisotropy possibly produced in response to stretching of the cuticle during expansion growth. Wax platelet patterns appear to be restricted to certain taxonomic entities; thus, genetical determinants are also involved in their formation.     

Fundamental Causes of the Global Patterns of Species Range and Richness1

Global patterns of species range and richness are a consequence of many interacting factors, including environmental conditions, competition, geographical area, and historical/evolutionary development. Two widely studied global patterns of distribution are the latitudinal and elevation gradients of species range and richness. The fundamental mechanisms by which environment and physiology of the plants themselves interact to generate global-scale correlations between increased species range or decreased species richness and latitude/elevation have not previously been established. This paper develops the hypothesis that the primary climatic variables determining global-scale gradients in ectotherm species range and richness are temperature (T) and temperature variability (T), and that the primary physiological variable defining adaptation of ectotherms to temperature is respiratory energy metabolism. This hypothesis is based on a postulate that adaptation of ectotherms to latitudinal/altitudinal gradients of T and T leads to corresponding gradients in properties of energy metabolism. The gradients of metabolic properties give rise to gradients of species range and richness that are observed on a global scale. We demonstrate that natural selection results in ectotherms with metabolic properties matched to their environment and that energy use efficiency and the temperature range allowing growth are inversely related. Thus, opposing selective pressures to increase metabolic energy-use-efficiency or to increase the probability of surviving climate extremes control adaptation of ectotherms to climate. The principles developed in this paper yield fundamental laws of ecology that allow calculation of the contributions of global temperature patterns to the formation of gradients of species range and diversity. Relative values of richness and range are calculated solely from data on abiotic variables. Predictions agree with known patterns of ectotherm distribution.     

Chemo-adoptive immunotherapy of nude mice implanted with human colorectal carcinoma and melanoma cell lines

Summary The antitumor effects of chemotherapy, recombinant human interleukin-2 (IL-2), recombinant human interferon A/D (IFN), allogeneic human lymphokine-activated killer (LAK) cells, and antitumor monoclonal antibody (mAb), administered alone and in various combinations, were tested in athymic nude mice carrying human tumor xenografts. Treatment began 6–18 days after i.v. or i.p. inoculation of colorectal carcinoma or melanoma cell lines, when macroscopic growths were evident. Chemotherapy consisted of two or three courses of 5-fluorouracil (5-FU) or dacarbazine. IL-2 and/or IFN were administered three to five times weekly for 1–3 weeks, usually starting 2–5 days after chemotherapy. Human LAK cells were infused once or twice weekly for 2 or 3 weeks concurrently with IL-2. In some experiments, murine anticolorectal carcinoma mAb (SF25) was administered. In both tumor systems, chemotherapy alone or immunotherapy alone (IL-2, IL-2 + LAK cells, IFN, IL-2 + IFN ± LAK cells) had little or no therapeutic effects. Additive effects were obtained by combining chemotherapy with IL-2 and LAK cells or with IL-2 and IFN. In the majority of the experiments, the most effective combination was chemotherapy + IL-2 + IFN + LAK cells. Treatment with mAb was beneficial in the colorectal carcinoma system when combined with 5-FU + IL-2 or 5-FU + IL-2 + IFN. Homing experiments with radiolabeled human and mouse LAK cells injected i.v. showed increased early accumulation in the liver and lungs, whereas freshly explanted mouse splenocytes localized mostly in the spleen and liver. The tissue distribution pattern of human LAK cells was similar in normal and tumor-bearing mice (with lung metastases). These findings suggest that combination of chemotherapy with cytokines and LAK cells can be partially effective for advanced solid human tumors even in the absence of the host's T-cell immune response. Preliminary experiments showed that tumor-specific, anti-melanoma T-cell clones were effective in local (s.c.) tumor growth inhibition (Winn assay) following coinjection with the autologous tumor cells.     

The control of pacemaker modulations for social communication in the weakly electric fish Sternopygus

Summary Nearly sinusoidal electric organ discharges (EODs) of the weakly electric fish Sternopygus, occur at a regular rate within a range from 50 to 200 Hz and are commanded by a medullary pacemaker nucleus (Pn). During courtship and aggression, the rate of EODs is modulated as smooth EOD-frequency rises or brief EOD-interruptions (Hopkins 1974b). The present study examines the control of such modulations. Rises were elicited by L-glutamate stimulation of the diencephalic prepacemaker nucleus, the only previously known source of input to the Pn. We demonstrate an additional input to the Pn, the sublemniscal prepacemaker nucleus (SPPn). L-glutamate stimulation of this area caused EOD-interruptions.The Pn contains electrotonically coupled pacemaker cells which generate the rhythm of the EODs, as well as relay cells which transmit the command pulse to the spinal motor neurons that innervate the electric organ. Pacemaker cells recorded intracellularly during EOD-interruptions continued firing at their regular frequency but with slightly increased jitter. Relay cells, on the other hand, were strongly depolarized and fired spikelets at a greatly increased frequency during EOD-interruptions. Thus EOD-interruptions were caused by SPPn input to relay cells that caused their massive depolarization, blocking the normal input from pacemaker cells without greatly affecting pacemaker cell firing characteristics.Application to the Pn of an antagonist to NMDA-type glutamate receptors blocked EOD-frequency rises and EOD-interruptions. Antagonists to quisqualate/ kainate receptor-types were ineffective.Abbreviations EOD Electric Organ Discharge - JAR Jamming Avoidance Response - Pn pacemaker nucleus - PPn diencephalic prepacemaker nucleus - SPPn sublemniscal prepacemaker nucleus     

Enzyme distribution patterns in the imaginal wing disc ofDrosophila melanogaster and other diptera

Summary Analysis of the development of the aldehyde oxidase (AO) pattern in the wing pouch ofD. melanogaster showed that the extension of areas with AO activity occurs in steps. This indicates that the activation of this enzyme is regulated in groups of cells. It is proposed to use the term territory for such a cell group. In the wing pouches ofD. melanogaster, D. simulans andMusca, corresponding parts of the disc become AO positive at comparable developmental stages. This indicates that AO becomes active in individual territories in a specific sequence.Borderlines of the distribution pattern of different enzymes in the wing pouch ofDrosophila and other dipteran species are in agreement with those found for the development of the AO pattern or are complementary to them. This indicates the existence of a common set of territories in the wing pouches of all higher diptera. Borderlines of patterns, as caused by different genetic constitution, are also in accord with this set of territories. The borderlines of some territories coincide with the compartmental A/P or D/V boundary. The results support the idea that both the location of compartmental boundaries and that of borderlines of enzyme territories are determined by a single mechanism.The distribution and the shape of the territories in the wing pouch is best explained by the reaction-diffusion model proposed by Meinhardt (1980), which involves three different gradients.     

The role of learning in fish behaviour

Summary The behavioural patterns of fish are the result of innate (built-in) patterns of maturation (developmental changes) and of learning processes (imprinting and trial-and-error learning). Innate behavioural patterns are considered to be hard-wired and inflexible. However, through learning, fish can adapt to environmental change. For instance, the homing behaviour of fish may be partly the result of the development of specific parts of the brain and partly because of changes in behaviour with experience. Similarly, one can assume that the feeding mode of fish involving snap-responses is innate, but learning enables fish to modify their foraging behaviour in response to a fluctuating environment. By reviewing these and other examples, such as the role of recognition learning and socially transmitted behaviour, one can illustrate the importance of learning in the everyday life of fishes. Although learning plays a large role in the behaviour of fishes, the learning capacity of fishes may also be useful to fisheries research and hatchery operations.     

Metatarsal robusticity in primates and a few other plantigrade mammals

By using a new technique for determining relative metatarsal robusticity, the distribution of the 1+5 pattern (in which M5 is the second most robust metatarsal after M1) and 15 pattern (in which M5 is the least robust of all metatarsals) was established in primates and a few other plantigrade mammals. The first pattern is associated with a terrestrial and the second with an arboreal substrate. Robusticity formulae are not connected with specific locomotor patterns, but a total robusticity quotient is associated with these patterns and substrate preference as well. Changes in substrate preference are accompanied by changes of total robusticity, an increased number of permutations and ultimately a change of the robusticity pattern. Intermetatarsal robusticity gradients are related to the direction and intensity of muscular activity. A combined analysis of all factors can reveal a great deal of the locomotor history of a taxon.Also ofThe Institute of Applied Biology, New York.     

Establishing and maintaining axial growth: wall mechanical properties and the cytoskeleton

Organ morphology depends on cell placement and directional cell expansion. Microtubules are involved in both of these processes so genetic approaches to understand the role microtubules play in organ expansion are not straightforward. Our use of the temperature-sensitive mor1-1 mutants led to the surprising discovery that Arabidopsis thaliana (L.) Heynh. root cells can establish and maintain transverse cellulose texture without well organized microtubule arrays. This work also demonstrated that cells can lose the ability to expand anisotropically without losing transversely oriented cellulose microfibrils. We suggest that microtubule disruption affects the cells ability to generate long cellulose microfibrils, which may be essential for achieving growth anisotropy. Thus organ shape may depend not only on the orientation but also on the relative length of cellulose microfibrils during axis establishment and growth. More recent work has shown an important correlation between microtubule organization and the deposition patterns of the glycosylphosphatidylinositol (GPI)-anchored wall protein COBRA. Loss of microtubule organization is associated with the dissipation of transverse banding patterns of COBRA, suggesting that COBRAs function in maintaining anisotropic expansion may be microtubule-dependent.     

Shaping Morphogen Gradients by Proteoglycans

During development, secreted morphogens such as Wnt, Hedgehog (Hh), and BMP emit from their producing cells in a morphogenetic field, and specify different cell fates in a direct concentration-dependent manner. Understanding how morphogens form their concentration gradients to pattern tissues has been a central issue in developmental biology. Various experimental studies from Drosophila have led to several models to explain the formation of morphogen gradients. Over the past decade, one of the main findings in this field is the characterization of heparan sulfate proteoglycan (HSPG) as an essential regulator for morphogen gradient formation. Genetic and cell biological studies have showed that HSPGs can regulate morphogen activities at various steps including control of morphogen movement, signaling, and intracellular trafficking. Here, we review these data, highlighting recent findings that reveal mechanistic roles of HSPGs in controlling morphogen gradient formation.Embryonic development involves many spatial and temporal patterns of cell and tissue organization. These patterning processes are controlled by gradients of morphogens, the “form-generating substances” (Tabata and Takei 2004; Lander 2007). Secreted morphogen molecules, including members of Wnt, Hedgehog (Hh), and transforming growth factor-β (TGF-β) families, are generated from organizing centers and form concentration gradients to specify distinct cell fates in a concentration-dependent manner. Understanding how morphogen gradients are established during development has been a central question in developmental biology. Over the past decade, studies in both Drosophila and vertebrates have yielded important insights in this field. One of the important findings is the characterization of heparan sulfate proteoglycan (HSPG) as an essential regulator for morphogen gradient formation. In this review, we first discuss various models for morphogen movement. Then, we focus on the functions of HSPGs in morphogen movement, signaling, and trafficking.     

Effect of light-dark cycles with different LD time-ratios and different LD intensity-ratios on the activity rhythm of chaffinches

Summary Chaffinches (Fringilla coelebs L.) were kept at light-dark (LD) cycles with two different light intensity ratios (ca. 500.1 and ca. 101 lux) both with 1.83 hours of twilight at the onset and end of light time. The light time/dark time ratio (LD time-ratio) was varied in the range from 420 to 222 hours.Birds at the high LD intensity-ratio (500.1 lux) increased activity time () with increasing light time from LD 420 to 222. Time of activity onset, middle, and end of activity corresponded to onset, middle, and end of light time from LD 420 to 168, but the phase angles of middle and end of activity (related to midpoint of light time and midpoint of artificial dusk respectively) became more positive (advanced) at LD 204 and 222.Birds at the lower LD intensity-ratio (101 lux) increased activity time with increasing light time from LD 816 to 204 but activity time at LD 420 was equivalent to that at LD 816. The phase angles of activity onset and middle (related to midpoint of artificial dawn and midpoint of light time, respectively) became more positive at both long (LD 204) and short (LD 420) light times.By use of different twilight regimes (linear and logarithmic changes in intensity), it was shown that end of activity was more closely related with the rate of change and/or absolute level of light intensity than was onset of activity.During January and February, the birds responded to increased LD time-ratios by an increase of activity time and by strong positive phase angle shifts which correspond to observations of activity patterns of chaffinches exposed to outdoor conditions during the spring breeding season.The free-running circadian periods () of 6 birds measured at 15 lux at the end of the experiments following LD 204 averaged 22.17 hours.From the data on precision, i. e. the period-to-period variation in the phase angles of single birds averaged over all birds of one group and condition, as well as from the number of birds not synchronized with the LD cycles, it is concluded that extreme long and short light times per 24 h represent weaker zeitgebers compared to LD cycles with medium lengths of light time. Likewise, LD cycles with low LD intensity-ratios are weaker zeitgebers than LD cycles with high LD intensity-ratios.This study was carried out while the senior author was on sabbatical leave from the University of Alaska and held a fellowship from the Alexander von Humboldt Stiftung.     

Exposure of Petunia Seedlings to Ethylene Decreased Apical Dominance by Reducing the Ratio of Auxin to Cytokinin

Seedlings of Petunia x hybrida Orchid treated with the ethylene-releasing compound ethephon at 0.9, 1.7, and 3.5 mM evolved ethylene at a higher rate as the concentration of ethephon increased. Regardless of the concentration of ethephon applied, ethylene evolution peaked 6 to 8 h following application. Evidence that ethephon application decreased apical dominance included an increase in the number of new nodes on the main stem and a sustained increase in the length of new and existing lateral shoots compared to the control (no ethephon). Plants treated with 3.5 mM ethephon developed mild chlorosis, whereas a concentration of 1.7 mM ethephon decreased apical dominance without phytotoxic effects. The auxin/cytokinin ratio decreased in the apical shoot section as early as 1 h after ethephon treatment. In contrast, a decrease in the ratio in the subapical shoot section was not detected until 24 h after ethephon application. Reduction in auxin/cytokinin ratio was a result of a decrease in indole-3-acetic acid (IAA) and an increase of zeatin riboside (ZR), but not isopentenyladenosine (iPA). These results suggest that exposing Orchid petunia seedlings to ethylene via ethephon lowers the auxin/cytokinin ratio, thereby promoting the outgrowth of lateral shoots.     

Looking back on the discovery of α-bungarotoxin

This review is a personal narration by a retiring pharmacologist from Taiwan who looks back at his discovery of -bungarotoxin from the historical perspective of Taiwan during the last 50 years, with accounts of his experiences and his efforts to overcome hardship. How the -toxin was isolated and characterized as an irreversible specific nicotinic acetylcholine (ACh) receptor antagonist,and how it subsequently became a useful experimental probe are presented here. The dilemma of differentiating the actions of tubocurarine and -bungarotoxin is analyzed. The author also outlines findings based on work done in his laboratory using -bungarotoxin as a tool on particular aspects of synaptic transmission. These include presynaptic receptor for positive feedback of transmitter release, explosive release of ACh, up- and down-regulation of ACh receptors after chronic drug treatment, autodesensitization of junctional ACh receptors, differences in action between natural transmitter and exogenous agonists and that between junctional and extra-junctional ACh receptors. Some experimental pitfalls, in which biomedical scientists are frequently trapped, are raised. Finally, some anecdotes are appended from which the reader may further understand scientific life in the 20th century, including its joys and regrets.     

Vergleichende Untersuchungen über die Insulinempfindlichkeit und den Einfluss von „Ganglienblockade“ auf den Insulineffekt

Zusammenfassung In vergleichenden Untersuchungen an verschiedenen Haustieren (Pferd, Schwein, Kaninchen, Schaf und Huhn) wird die unterschiedliche Insulinresistenz geprüft. Insulinempfindlich sind Pferd, Schwein und Kaninchen, während sich Schaf und Huhn als weitgehend insulinresistent erweisen.Durch gleichzeitige Verabreichung von Insulin und N,N,N,N-3-pentamethyl-N,N-diäthyl-3-aza-pentan-1,5-diammonium-dibromid (P) gelingt es, bei allen untersuchten Species die Insulinhypoglykämie zu verstärken.Die Verstärkung der Insulinhypoglykämie durch P wird als Hemmung der gegenregulatorischen neurogenen Adrenalinausschüttung gedeutet.Tierartliche Besonderheiten in der Blutzuckerregulation werden diskutiert.     

Kinetics of the flash-induced electrochromic absorbance change in the presence of background illumination. Turnover rate of the electron transport. II. Higher plant leaves

The flash-induced electrochromic absorbance change (A ₅₁₅) was measured in leaves of higher plants in the absence and presence of continuous monochromatic background illumination of different intensities and wavelengths. The variation of the amplitude of A ₅₁₅ in background light was used to estimate the steady-state turnover time of the electron transport. In red light we obtained about 5 msec which was accounted for by the turnover of the linear electron transport. With far red background illumination or in the presence of the photosystem 2 inhibitor, DCMU, the steady-state turnover time tentatively assigned to photosystem 1 cyclic electron transport was much larger (100 msec).Increasing the intensity of background illumination with far red light gradually diminished the slow rise of A ₅₁₅ in parallel with suppression of the initial rise generated by photosystem 1. At high intensities of the red light, however, while A ₅₁₅ was attenuated, the slow rise was not eliminated and its proportion relative to the initial rise did not vary appreciably.     

Hoxd10 induction and regionalization in the developing lumbosacral spinal cord

We have used Hoxd10 expression as a primary marker of the lumbosacral region to examine the early programming of regional characteristics within the posterior spinal cord of the chick embryo. Hoxd10 is uniquely expressed at a high level in the lumbosacral cord, from the earliest stages of motor column formation through stages of motoneuron axon outgrowth. To define the time period when this gene pattern is determined, we assessed Hoxd10 expression after transposition of lumbosacral and thoracic segments at early neural tube stages. We present evidence that there is an early prepattern for Hoxd10 expression in the lumbosacral neural tube; a prepattern that is established at or before stages of neural tube closure. Cells within more posterior lumbosacral segments have a greater ability to develop high level Hoxd10 expression than the most anterior lumbosacral segments or thoracic segments. During subsequent neural tube stages, this prepattern is amplified and stabilized by environmental signals such that all lumbosacral segments acquire the ability to develop high levels of Hoxd10, independent of their axial environment. Results from experiments in which posterior neural segments and/or paraxial mesoderm segments were placed at different axial levels suggest that signals setting Hoxd10 expression form a decreasing posterior-to-anterior gradient. Our experiments do not, however, implicate adjacent paraxial mesoderm as the only source of graded signals. We suggest, instead, that signals from more posterior embryonic regions influence Hoxd10 expression after the early establishment of a regional prepattern. Concurrent analyses of patterns of LIM proteins and motor column organization after experimental surgeries suggest that the programming of these characteristics follows similar rules.