Analysis of the secretions of the subcommissural organs of several vertebrate species by use of fluorescent lectins

Summary The glycoprotein secretions of the subcommissural organ were analyzed with the use of nine fluorescent lectins, specific to different sugar moieties. After exposure to Concanavalin A a bright fluorescence was observed in the ependymal cells of the subcommissural organs of all vertebrates studied (Lampetra planeri, Ameiurus nebulosus, Bufo bufo, Lacerta vivipara, Gallus gallus, Rattus norvegicus, Ovis aries). The fluorescence is abolished by the competitive sugar, -D-mannopyranosyl. The intensity of the lectin fluorescence decreases from the phylogenetically lower to the higher forms, paralleled by a change in polarity of the secretion from a vascular (lower vertebrates) to a ventricular (higher vertebrates) direction. The strong affinity for Concanavalin A suggests the presence of a glycoprotein rich in mannosyl residues in the ependymal cells and a similarity of composition of this glycoprotein among the vertebrates. Lens culinaris agglutinin and wheat germ agglutinin revealed fluorescent rosettes in the hypendymal cells of the sheep. Binding of both these lectins suggests the presence of a glycoprotein rich in N-acetyl-D-glucosamine.In the underlying ventricular cavity, no fluorescence could be observed, suggesting that the Reissner's fiber does not possess the same carbohydrate constitution as the ependymal secretion of the subcommissural organ.     

Cytokine production in whole blood cell cultures of patients undergoing therapy with biological response modifiers or 5-fluorouracil

We have measured the levels of interferon (IFN), tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-1, and IL-2 in the whole blood cell culture supernatants of 43 tumor patients undergoing a treatment with biological response modifiers or a conventional therapy with 5-fluorouracil and leucovorin. In the blood cell cultures of the 16 patients who received 5-fluorouracil and leucovorin IFN levels decreased (P0.01) and TNF levels rose (P0.05) during each therapy cycle. However, in the blood samples a declining number of total leukocytes and lymphocytes was measured (P0.05). Progressive disease could be correlated to a tendency towards lower IFN levels in the pretherapeutic cultures of these patients. The second group analyzed consisted of 8 patients receiving a low-dose IL-1 therapy. In this group we found either an unchanged or an augmented IFN production of the blood cells during treatment. In the group of 13 patients receiving low-dose recombinant IL-2 (4.5×10⁶IU m^–2 day^–1) significantly increasing IFN levels were seen in the blood cell cultures during the therapy (P0.05), although total leukocyte counts decreased. In this group, 4 had stable disease for at least 2 months and 9 patients had tumor progression under therapy. In the cultures of the latter a tendency towards lower IFN values was found. Finally, the cytokine production in the blood cell cultures of 6 patients receiving a combination therapy of IFN and high-dose IL-2 was studied. During this therapy a dramatically reduced production not only of IFN but also of all other measured cytokines was found. In this group all patients had tumor progression under therapy. It is concluded that the measurements of cytokine production in a reproducible whole blood culture system may be useful for monitoring immunological therapies and may help us to find out which doses of biological response modifiers have enhancing or suppressive effects on the functions of the immune cells.     

Role of homology and pathway specificity for recombination between plasmids and bacteriophage lambda

Summary To determine the minimum amount of homology required for efficient recombination in Escherichia coli, we measured recombination frequencies between bacteriophage and pBR322 derivatives containing DNA fragments of various sizes by assaying for phages that could transduce the bla and ori genes of pBR322. Efficient recombination required about 40 bp of homology; increases in homology above 40 bp resulted in proportionate increases in recombination, while decreases below 40 bp resulted in precipitous decreases in recombination. The recA ⁺ gene stimulated recombination over the entire range of homologies tested. Restriction enzyme digests of several recombinant DNA molecules indicated that they contained the complete plasmid DNA inserted in the genome as expected for a reciprocal crossover. Analysis of recombination frequencies in different recombination-deficient mutant strains indicated that the formation of -plasmid cointegrates by homologous recombination proceeded predominantly by the RecBC pathway and very inefficiently, if at all, by the RecE and RecF pathways.     

Lipase-catalyzed reactions of fatty acids with glycerol and acylglycerols

Summary Esterification and/or acidolysis reactions of oleic acid with glycerol, monooleoylglycerols, dioleoylglycerols, and trioleoylglycerol have been investigated using two immobilized microbial lipase preparations: Lipase G from Penicillium sp. and sn-1,3-specific Lipozyme from Mucor miehei. Lipase G effectively catalyzes the esterification of oleic acid with glycerol yielding monooleoylglycerols as the most predominant product, but it is rather ineffective for the esterification and/or acidolysis with the acylglycerols. Lipozyme also catalyzes the esterification of oleic acid with glycerol, although at a lower rate than Lipase G, and the predominant products formed are dioleoylglycerols, followed by monooleoylglycerols and trioleoylglycerol. With Lipozyme as biocatalyst, oleic acid reacts with monooleoylglycerols predominantly by esterification, with dioleoylglycerols to a greater extent by acidolysis than by esterification, and with trioleoylglycerol almost exclusively by acidolysis at the sn-1,3-positions. Potential applications of these lipase-catalyzed reactions in the industrial preparation of acylglycerols and deacidification of fats and oils by esterification of the fatty acids with endogenous acylglycerols or exogenous glycerol and/or acylglycerols are outlined.     

Callus proliferation from the protoplasts of embryogenic cells of Quercus serrata

Embryogenic culture was induced from the immature embryos of Quercus serrata using Marashige and Skoog's medium (MS) containing 0.1 M each of 2,4-d and BAP, and subcultured for seven months before isolation of protoplasts by using 1% Cellulase RS in 0.6 M mannitol solution. Efficient colony formation was obtained when protoplasts were cultured in a liquid MS medium containing 0.6 M mannitol, 3% sucrose and combination of 0.1 M or 1 M each of 2,4-d and BAP. Excluding ammonium nitrate from the MS medium resulted in the decrease of the percentage of colony formation. From colonies, both agar culture and liquid culture were sustained in the MS media without mannitol containing no plant growth regulator, or containing 0.1 M of BAP in combination with 0.1 M or 1 M of 2,4-d.Abbreviations BAP 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - MS medium after Murashige & Skoog (1962).     

Formation of the Interaction between the Wave Components of the Main EEG Rhythms in Children up to Five Years of Age

Using the analysis of the interaction between EEG components [1], the authors studied the regularities of the formation of the EEG wave structure in 36 children aged 4 to 7 months, 2 to 3 years, and 4 to 5 years. The EEG of 4- to 7-month-old children had a relatively organized temporal structure, whose components were connected mainly with those of the slow-wave range. This structure is reproduced in most of the leads and is more stable in the anterior cortical areas than in the posterior ones. The waking 2- to 5-year-old children had two functional nuclei in the relation structure of the EEG components characterized by statistically significant enhancement in the probability of the consecutive appearance of waves of certain ranges. These nuclei include a slow-wave nucleus in the range of the ₂–₁-frequencies and an alpha-nucleus in the ₁- and ₂-ranges (₂ and ₁ 2- to 3-year-old children) of the EEG frequencies. The relations between the components of the slow-wave nucleus begin to form in infants, prevail in 2- to 3-year-old children, and weaken by the age of 4 to 5 years. A trend is observed in 2- to 5-year-old children towards an increase in the average frequency of the waves that form the slow-wave nucleus. The temporal (strengthening interaction between the waves) and spatial organization of the interaction between the wave components of the alpha-nucleus is accelerated at the age of 4 to 5 years. The average frequency of the components forming this nucleus gradually increases with the children's age from 4 to 7 months to 4 to 5 years. Competitive relations exist between the two nuclei, characterized by a significant decrease in the probability of transitions between the components of different nuclei. The ₂-component may play a special role of the connecting link between the nuclei at the age of 4 to 5 years, the interwave transition from the - to -frequencies and from the latter to the -range being effected by a leaplike rather than smooth frequency increase. The character of local and spatial rearrangements of the EEG temporal structure detected in this study reflects a gradual morphofunctional maturation of the brain as a system that maintains the specific features of self-regulation mechanisms and coordination of the intersystem interactions at various stages of a child's development.     

Three-dimensional and quantitative observation of the hypertrophy of rat corticotrophs following adrenalectomy

Summary The three-dimensional form of corticotrophs in the anterior pitiutary gland of rats was studied by reconstruction from serial semi-thin sections both in control rats and rats one week after adrenalectomy. The corticotrophs have large depressions and cup-shaped cavities on their surface, and these features became more conspicuous after adrenalectomy. The hypertrophy of corticotrophs in adrenalectomized rats was quantified by measuring the area and perimeter of all serially sectioned profiles. The volume of the whole cell increased from 1129±114 m³ to 2902±201 m³ (P<0.01) after adrenalectomy, while the surface area of the cells increased from 690±45 m² to 1431±116 m² (P<0.01). The volume of the nucleus increased from 87±11 m³ to 172±14 m³ (P<0.05). Though the complexity of the form of corticotrophs seems to be increased after adrenalectomy, the ratios (adrenalectomized/control) of cell volume and surface area were 2.57 and 2.07, respectively; this indicates that the increase of the cell volume was greater than that of the surface area.     

Changes in EEG Power Spectra During Biofeedback of Slow Cortical Potentials in Epilepsy

The goal of the study was to explore parallel changes in EEG spectral frequencies during biofeedback of slow cortical potentials (SCPs) in epilepsy patients. Thirty-four patients with intractable focal epilepsy participated in 35 sessions of SCP self-regulation training. The spectral analysis was carried out for the EEG recorded at the same electrode site (Cz) that was used for SCP feedback. The most prominent effect was the increase in the 2 power (6.0–7.9 Hz) and the relative power decrement in all other frequency bands (particularly 1, 2, and 2) in transfer trials (i.e., where patients controlled their SCPs without continuous feedback) compared with feedback trials. In the second half of the training course (i.e., sessions 21–35) larger power values in the , , and bands were found when patients were required to produce positive versus negative SCP shifts. Both across-subject and across-session (within-subject) correlations between spectral EEG parameters, on the one hand, and SCP data, on the other hand, were low and inconsistent, contrary to high and stable correlations between different spectral variables. This fact, as well as the lack of considerable task-dependent effects during the first part of training, indicates that learned SCP shifts did not directly lead to the specific dynamics of the EEG power spectra. Rather, these dynamics were related to nonspecific changes in patients' brain state.     

Expectancy influence on self-reported experience during alpha feedback training

From a screening questionnaire, 12 subjects indicating some prior knowledge of alpha training (but no actual biofeedback training experience) and 12 subjects lacking such prior knowledge were selected for two 30-minute feedback sessions, one providing an opportunity to enhance and the other to suppress EEG alpha. Written self-reports of experiential state were independently rated by two judges on a five-point scale (Walsh, 1974) according to the degree to which they approximated alleged alpha vs. nonalpha experiences. These self-reports indicated that only the subjects expressing prior knowledge were able to differentiate between the enhancement and suppression sessions. A second study explored whether knowledgeable subjects (presumably those with positive expectancies) could make a similar differentiation when their expectancies regarding the type of feedback they would receive were manipulated. Twenty subjects received enhancement and 20 others received suppression feedback for four 40-minute sessions. Within each of these groups, 10 subjects were led to believe they enhanced alpha, and 10 were told they suppressed alpha. Half the subjects, therefore, believed they were attempting to change alpha in a direction opposite that of their actual task. Both actual and expected direction of alpha change significantly affected self-reports in the expected direction. Seemingly, situational expectancy alone was not sufficient to account for the actual feedback effects in the knowledgeable subjects. It is not entirely clear whether the alpha-related experiences reported by these knowledgeable subjects were responses to internal (i.e., physiologic state) or subtle external cues.     

Postembryonic development ofParalabidocera antarctica (I. C. Thompson) (Copepoda,Calanoida) from the fast ice near Syowa Station,Antarctica

Six nauplius and five copepodid stages as well as adults ofParalabidocera antarctica (I. C. Thompson, 1898) (Copepoda:Calanoida) are described based on specimens obtained from fast ice and collected by a plankton net near Syowa Station (69°00 S, 39° 35 E), Antarctica. The adult male and female are redescribed in detail. Nauplius stages ofP. antarctica are very similar to the previously describedAcartia species. Sexual dimorphism becomes apparent from copepodid IV onwards in the morphology of antennule and leg 5. The copepodid stages of this species retain certain characteristics not only of Acartiidae but also of Pontellidae and Parapontellidae.     

rDNA targeted oligonucleotide primers for the identification of pathogenic yeasts in a polymerase chain reaction

Summary Species-specific oligonucleotide primers were designed for PCR identification of the basidiomycetous yeastsCryptococcus neoformans, Trichosporon cutaneum andRhodotorula mucilaginosa. The procedure uses standard PCR components including DNA from the test species and three primers: two universal external (upstream and downstream) limiting primers and a species-specific internal primer. Species identification requires the formation of a species-specific rDNA nucleotide segment that is significantly smaller (200 bp) than a non-target segment (600 bp). The procedure can be used to identify yeasts from single and mixed populations.This paper is dedicated to Professor Herman Jan Phaff in honor of his 50 years of active research which still continues.     

Correlation between decreased reparative capability of UV lesions and higher spontaneous mutability in a mutator strain of E. coli K-12

Summary The biauxotrophic strain of E. coli K-12 (), met 1/his 7, which exhibits an 8 times higher rate of met1met ⁺-backmutation as compared with the parent strain met 1, was found to be also more sensitive to UV irradiation. In addition, the maximum UV induction of prophage occurs at lower doses, and the capacity of the strain to propagate induced prophage is reduced. The mutant strain has also lost part of the ability to repair UV-induced lesions in phage T 1.The phenotypes high mutability and UV sensitivity could not be separated by means of recombination.From these findings it is concluded that part of the spontaneously occuring changes in DNA (premutations) which lead to met ⁺ genotype are similarly repaired as some UV induced ones, and that the mutator mum ⁺ of strain met 1/his 7 causes a reduced repair of both changes.     

Microphotometric determination of enzyme activities in type-grouped fibres of reinnervated rat muscle

Summary Activities of succinate dehydrogenase (SDH), glycerolphosphate oxidase (GP-OX), cytochrome oxidase (CYT-OX) and lactate dehydrogenase (LDH) were determined microphotometrically in single, actomyosin-ATPase typed (Guth and Samaha 1970) fibres within cross-sections of normal and reinnervated rat tibialis anterior muscles. SDH and GP-OX activities displayed pronounced scattering and large overlaps existed between -, -, and -fibres of normal muscle. Coefficients of variation were in the range of 16–40% for GP-OX and SDH in the different fibre populations. Enzyme activity determinations in typegrouped -, -, and -fibres of reinnervated muscle showed much less scattering than in normally distributed -, -, and -fibres of control muscles. Coefficients of variation were in the range of 10–13% for SDH, GP-OX, CYT-OX and LDH. The experimental error of the kinetic microphotometric measurement of enzyme activities in situ is in the range of 10% (Reichmann and Pette 1982). Our results therefore suggest a high degree of metabolic similarity or homogeneity of typed-grouped muscle fibres and thus support the assumption that type-groped fibres are homogeneous and correspond to regularly assembled motor units.     

Ion channel hypothesis for Alzheimer amyloid peptide neurotoxicity

Summary 1. Alzheimer's disease (AD) is a chronic dementia and neurodegenerative disorder affecting the oldest portions of the population. Brains of AD patients accumulate large amount of the AP peptide in amyloid plaques.2. The AP[1–40] peptide is derived by proteolytic processing from a much larger amyloid precursor protein (APP), and has been circumstantially identified as the toxic principle causing cell damage in the disease.4. The AP[1–40] peptide is able to form quite characteristic calcium channels in planar lipid bilayers. These channels have conductances in the nS range, and can dissipate ion gradients quickly. The peptide can also cause equivalent cation conductances in cells.5. We suggest that amyloid channel blocking agents might be therapeutically useful in Alzheimer's Disease, and have constructed molecular models of the channels to aid in the design of such compounds.     

A New Method for Self-Regulation of Slow Cortical Potentials in a Timed Paradigm

A new method of slow cortical potential (SCP) biofeedback is described, in which subjects were presented with a sequence of two alternating tones. Subjects learned to adjust their SCPs with the 4-s rhythm of presented tones by producing directed SCP changes only in certain inter-tone intervals. Specifically, they learned to simultaneously produce two EEG signals: 1) positive or negative SCP shift at vertex, and 2) SCP asymmetry between the right and the left central area. After one training session, 13 healthy participants were able to differentiate significantly between the negativity and the positivity conditions; this differentiation was achieved within less than 300 ms after the discriminative signal, i.e. much faster than in previous studies employing traditional SCP biofeedback technique. However, these participants did not produce a significant hemispheric asymmetry in the first session. In the second experiment, five subjects participated in prolonged training (6 to 17 sessions). Highly significant control of SCP asymmetry over the precentral cortex was attained in four out of five participants. Advantages and disadvantages of the new method as compared with the classical SCP biofeedback technique are discussed.     

Charles Darwin's theory of evolution: A review of our present understanding

The paper characterizes Darwin's theory, providing a synthesis of recent historical investigations in this area. Darwin's reading of Malthus led him to appreciate the importance of population pressures, and subsequently of natural selection, with the help of the wedge metaphor. But, in itself, natural selection did not furnish an adequate account of the origin of species, for which a principle of divergence was needed. Initially, Darwin attributed this to geographical isolation, but later, following his work on barnacles which underscored the significance of variation, and arising from his work on botanical arithmetic, he supposed that diversity allowed more places to be occupied in a given region. So isolation was not regarded as essential. Large regions with intense competition, and with ample variation spread by blending, would facilitate speciation. The notion of place was different from niche, and it is questioned whether Darwin's views on ecology were as modern as is commonly supposed. Two notions of struggle are found in Darwin's theory; and three notions of variation. Criticisms of his theory led him to emphasize the importance of variation over a range of forms. Hence the theory was populational rather than typological. The theory required a Lamarckian notion of inheritable changes initiated by the environment as a source of variation. Also, Darwin deployed a use/habit theory; and the notion of sexual selection. Selection normally acted at the level of the individual, though kin selection was possible. Group selection was hinted at for man. Darwin's thinking (and also the exposition of his theory) was generally guided by the domestic-organism analogy, which satisfied his methodological requirement of a vera causa principle.     

The new problem of genetics: A response to Gifford

Recently, Fred Gifford attempted to explicate the meaning of the term genetic as applied to phenotypic traits. He takes as his primary goal the explication of how the term is used and tries to avoid conclusions about how it should be used. He proposes two independent criteria (DF and PI) which together capture much of what biologists mean when they describe traits as genetic. Although Gifford's approach is extremely insightful in many ways, I argue that his analysis is not sufficiently critical concerning the adequacy of common usage.In particular, while DF is a perfectly legitimate and useful measure of heritability in populations, it is not necessarily a genetic one and should not be labeled as such. PI on the other hand, although very intuitive, depends on an extremely problematic distinction between causes and mere conditions (e.g., genes and epigenetic factors). Both criteria will be highly relative and both, via what I term the new problem of genetics, will inspire contradictory analyses based on the same data.Fortunately, as Gifford recognizes, it is not necessary to make sense of genetic at all in order to do biology. Quantitative genetics can do the kind of (heritability) analysis that DF embodies without making questionable claims about genes. Causal-mechanical or bottom-up biology can proceed perfectly well without postulating the priveleged role for genetic causes that PI entails. In short, talk of genetic traits, under either criteria, is unnecessary and misleading.     

Comparisons of δ13C values in leaves of aquatic macrophytes from different habitats in Britain and Finland; some implications for photosynthetic processes in aquatic plants

Summary The ¹³C values of submerged aquatic plants from contrasting but relatively defined habitats, and the ¹³C values of emergent, floating and submerged leaves of dimorphic aquatic plants, were measured. In many instances the ¹³C values of dissolved inorganic carbon in the water were also measured. Plant ¹³C values in the vicinity of-40 to-50 were found in rapidly flowing spring waters with carbonate ¹³C values of-16 to-21, consistent with the notion that species such as Fontinalis antipyretica almost exclusively assimilate free CO₂ via RuP₂ carboxylase. Plant ¹³C values in the vicinity of-10 to-15 in sluggish water with carbonate ¹³C values of about-5 were observed, consistent with the notion that boundary layer diffusion and/or HCO₃ ^- uptake may determine the ¹³C value of submerged aquatic plants in these circumstances. Comparisons of ¹³C values of the same or related species growing in waters of similar carbonate ¹³C value but different flow rates confirmed this view; more negative ¹³C values were frequently associated with plants in fast moving water. In Britain, but not in Finland, the ¹³C values of submerged leaves of dimorphic plants were almost invariably more negative than in aerial leaves. The ¹³C value of carbonate from chalk streams and in acid springs indicate substantial inputs of respiratory CO₂, as opposed to atmospheric carbon. The contributions of these variations in ¹³C of the carbon source, and of isotope fractionation in diffusion, to the ¹³C value of submerged parts of dimorphic plants is discussed.     

Subtle behavioral variation in wild chimpanzees, with special reference to Imanishi’s concept of kaluchua

Here we consider the concept of kaluchua (a word adopted from the English culture) in group-living animals developed by Imanishi in the 1950s. He distinguished it from bunka (the Japanese equivalent to the English culture) because he thought that bunka had strong connotations of noble and intellectual human-like activities. Although he did not rigidly define kaluchua, his original concept of kaluchua was much broader than bunka and represented non-hereditary, acquired behavior that was acknowledged socially. However, instead of social life, complex feeding skills have often formed the central topic in the current studies of animal culture. In order to provide evidence that more subtle behavioral variations exist among wild chimpanzee (Pan troglodytes) populations, we directly compared the behaviors of two well-habituated chimpanzee groups, at Bossou and Mahale. During a 2-month stay at Bossou, M.N. (the first author) saw several behavioral patterns that were absent or rare at Mahale. Two of them, mutual genital touch and heel tap were probably customary for mature females and for mature males, respectively. Index to palm and sputter are still open to question. These subtle patterns occurred more often than tool use during the study period, suggesting that rarity is not the main reason for their being ignored. Unlike tool use, some cultural behavioral patterns do not seem to require complex skills or intellectual processes, and sometimes it is hard to explain the existence of such behaviors only in terms of function.