Genetic and biochemical studies on perchloroethylene ‘in vitro’ and ‘in vivo’

Perchloroethylene (PCE) was tested in a diploid strain (D7) of the yeast Saccharomyces cerevisiae in suspension tests with and without a mammalian microsomal activation system (S9) and ‘in vivo’ by the intrasanguineous host-mediated assay. In addition, enzyme alteration studies were performed in mice non-pretreated or pretreated with phenobarbital + β-naphthoflavone. PCE did not induce any genetic effect either ‘in vitro’ or ‘in vivo’. In the suspension test, PCE was more toxic without metabolic activation and less toxic with mammalian microsomal activation. The enzymatic determinations showed an increase of the aminopyrine demethylase activity and of the level of cytochrome P-450.     

Scanning electron microscopy of pollen structure throws light on resolving Bambusa–Dendrocalamus complex: bamboo flowering evidence

Since the introduction of the new genus Sinocalamus in 1940 which is now dissociated into Bambusa and Dendrocalamus, molecular markers have long been unable to discern members of the Bambusa–Dendrocalamus complex. Rapid concerted evolution governed by high level of transition/transversion at the noncoding DNA regions has limited the ability of internal transcribed spacer (rDNA) and trnL-F intergenic spacer to resolve Bambusa and Dendrocalamus in phylogenetic analysis. Based on scanning electron microscopy (SEM) analysis of Bambusa vulgaris and Dendrocalamus manipureanus pollen development, we provided the evidence that there exists genus specificity in the development and structure of woody bamboo pollen which can serve as a benchmark for allocated new species into the genus Bambusa and Dendrocalamus substantiating molecular data.     

The effect of cytokinins on shoot proliferation,biochemical changes and genetic stability of Rhododendron ‘Kazimierz Odnowiciel’ in the in vitro cultures

Plant Cell, Tissue and Organ Culture (PCTOC) - Shoot proliferation is a very important micropropagation phase, decisive for economic efficiency of this method for a given taxon. To obtain a high...     

Oddity and the ‘confusion effect’ in predation

We report on two sets of experiments designed to clarify the roles of sensory ‘confusion’ and prey ‘oddity’ as they interact to influence the hunting success of a pursuit predator, the largemouth bass (Micropterus salmoides), on silvery minnows (Hybognathus nuchalis). Bass quickly captured solitary minnows, but performed many unsuccessful attacks and took much longer to make a capture as prey school size was increased. At school sizes of eight and above, bass were effectively stymied, demonstrating the ‘confusion effect’. The inclusion of one or two ‘odd’ (blue-dyed) minnows in a school of eight greatly increased the ability of bass to capture both normal and odd prey, but this effect of oddity disappeared at a school size of 15. The implications of these results for understanding the adaptive basis of mixed species flocks, herds and schools is discussed.     

Neuroprotective effect of the hexapeptide HLDF-6 on rat hippocampal neurons on the in vivo and in vitro models of alzheimer’s disease

The neuroprotective effect of Thr-Gly-Glu-Asn-His-Arg hexapeptide (HLDF-6), a biologically active fragment of the differentiation factor of human leukemia cells (HLDF), was demonstrated on models of Alzheimer’s disease in vivo and in vitro. The syndromes of this pathology were induced in male rats by injection of beta-amyloid peptide (25–35) and ibotenic acid into the hippocampus. HLDF-6 prevented loss of long-term memory and decrease in the exploratory behavior of these animals and significantly decreased the number of pyknotic neurons in the CA1 area of the hippocampus. This peptide also exerts a protective effect in vitro on the primary cultures of the rat hippocampal and cerebellar neurons under conditions of the beta-amyloid toxicity. An increase in the dihydrotestosterone (DHT) content was demonstrated in the blood plasma of rats with the syndrome of Alzheimer’s disease and in the medium of the culture of hippocampal neurons in the presence of the Aβ(25–35) peptide. HLDF-6 inhibited this increase in both cases. A probable mechanism of the neuroprotective effect of HLDF-6 was suggested as being connected to its possible effect on both the biosynthesis and the metabolism of sex steroid hormones.     

In?vitro and in?vivo study on the effect of antifungal agents on hematopoietic cells in mice

Liposomal amphotericin B, voriconazole, and caspofungin are currently used for systemic and severe fungal infections. Patients with malignant diseases are treated with granulocyte-colony stimulating factor (G-CSF) for the recovery of granulocytes after chemotherapy or hematopoietic cell (HC) transplantation. Since they have a high incidence of fungal infections, they inevitably receive antifungal drugs for treatment and prophylaxis. Despite their proven less toxicity for various cell types comparatively with amphotericin B and the decrease in the number of leukocytes that has been reported as a possible complication in clinical studies, the effect of liposomal amphotericin B, voriconazole, and caspofungin on HCs has not been clarified. The present study aimed to examine the in vitro and in vivo effect of these three modern antifungals on HCs. Colony-forming unit (CFU) assays of murine bone marrow cells were performed in methylcellulose medium with or without cytokines and in the presence or absence of various concentrations of liposomal amphotericin B, voriconazole, and caspofungin. In the in vivo experiments, the absolute number of granulocytes was determined during leukocyte recovery in sublethally irradiated mice receiving each antifungal agent separately, with or without G-CSF. In vitro, all three antifungal drugs were nontoxic and, interestingly, they significantly increased the number of CFU-granulocyte-macrophage colonies in the presence of cytokines, at all concentrations tested. This was contrary to the concentration-dependent toxicity and the significant decrease caused by conventional amphotericin B. In vivo, the number of granulocytes was significantly higher with caspofungin plus G-CSF treatment, higher and to a lesser extent higher, but not statistically significantly, with voriconazole plus G-CSF and liposomal amphotericin B plus G-CSF treatments, respectively, as compared with G-CSF alone. These data indicate a potential synergistic effect of these antifungals with the cytokines, in vitro and in vivo, with subsequent positive effect on hematopoiesis.     

The effect of antioxidants on in vivo and in vitro methemoglobin formation in erythrocytes of patients with Parkinson’s disease

Methemoglobin formation was examined in erythrocytes of 16 patients with Parkinson’s disease (PD) (stage 3–4 by the Hoehn and Yahr scale). The patients receiving levodopa-containing drugs (madopar, nakom) were also treated with intramuscular injections of mexidol (daily dose 100 mg/day) for 14 days. Control group included 12 clinically healthy persons. The erythrocyte methemoglobin content was determined by electronic paramagnetic resonance (EPR) using the EPR signal intensity with the g-factor 6.0. The methemoglobin content was significantly higher in erythrocytes of PD patients than in healthy donors. The complex therapy with mexidol normalized the methemoglobin content in erythrocytes of PD patients. Incubation in vitro of erythrocytes of donors and PD patients with acrolein increased the methemoglobin content, while incubation with carnosine normalized the methemoglobin content in erythrocytes of PD patients. Prophylactic (i.e. before acrolein addition) and therapeutic administration of carnosine to the incubation system with acrolein decreased the methemoglobin content to its initial level. Results of this study suggest that inclusion of the antioxidants mexidol and carnosine in the scheme of basic therapy of PD may reduce side effects associated with methemoglobinemia.     

Radiocaesium in an organic soil and the effect of treatment with the fungicide ‘Captan’

Soil fungi accumulate radiocaesium from contaminated soil and it has been hypothesised that this may alter the plant availability and movement of the radionuclide in soil. The effect of twice-monthly addition of an aqueous suspension of the fungicide ‘Captan’ on the changes in a peaty podzol soil at 2 sites, contaminated 2 or 3 years earlier by the injection of ¹³⁴Cs, has been quantified. The sites had different soil acidity and vegetation cover. The less acid soil (pH_water 5.0) had been improved by the addition of lime and fertilizer and was reseeded with grass and clover. The more acid soil (pH_water 3.8) was under hill grasses, herbs and heather. On both sites the addition of fungicide did not alter the amount or concentration of radiocaesium in plant material sampled monthly or the depth distribution of radiocaesium in the soil profile. The concentration of the fungal constituent, ergosterol, in the soil, measured monthly, was unaffected by the fungicide treatment but evidence was obtained from a pot experiment to show that ergosterol decomposes slowly in cold, wet soils. On the more acid soil, two weeks after the last application of fungicide, there was a decline in active fungi as measured by fluorescein diacetate staining. Chloroform fumigation of the more acid soil resulted in a small increase in the amount of ¹³⁴Cs exchangeable with 1 M ammonium acetate. Radiocaesium in seven different fungi grown in pure culture was found to be almost entirely extractable (> 95%) with 1 M ammonium acetate. Another, Amanita rubescens, showed some retention and 88% was extractable. These findings do not preclude the fungal biomass as an important soil component controlling plant availability of radiocaesium from acid, organic soils by maintaining radiocaesium in a biological cycle, but make it unlikely that any fixation by fungi in a chemical sense is involved.     

In vitro production of flowering shoots in ‘German Red’ carnation: effect of uniconazole and gibberellic acid

Summary Callus regenerated near the base of senescing petals of flower bud explants of German Red carnation (Dianthus caryophyllus L.) produced adventitious flowering microshoots on MS-medium containing benzylaminopurine (8.9 M) and naphthaleneacetic acid (2.7 M). When these microshoots were subcultured with some callus, additional adventitious flowering microshoots were produced from the callus. The production of adventitious flowering shoots continued for many subcultures spanning a period of more than two years. Uniconazole (6.9 M) increased the number of adventitious shoots formed by as much as two-fold but decreased shoot length by about 50%. In contrast, GA₃ (2.9 M) decreased adventitious shoot formation and increased shoot length. Regardless of the growth regulator treatment, virtually all of the adventitious shoots produced flower buds. Thus, the growth regulators influenced flowering only indirectly by altering the number of adventitious shoots produced in vitro. These results demonstrate that the flowering habit of the adventitious shoots of German Red carnation is highly persistent and the flowering stimulus continues to be transmitted to the newly formed microshoots through the callus.     

‘In vitro’ and ‘in situ’ decomposition of nuisance macroalgae Cladophora glomerata and Pilayella littoralis

The decomposition of two macroalgal species Cladophora glomerata (CHLOROPHYTA) and Pilayella littoralis (PHAEOPHYTA) was studied in the laboratory and field conditions. These species are known to cause the extensive macroalgal blooms in the whole coastal range of the Baltic Sea. The objective of the experiments was to determine decomposition rates of the macroalgae, follow the changes in tissue nutrient content and validate the role of benthic invertebrates in this process. In the laboratory conditions, the differences in the decomposition rates of the algae were mainly due to the oxygen conditions. The weight loss of C. glomerata was slightly higher in anaerobic conditions than in aerobic conditions. If 99% of initial dry weight of P. littoralis was lost in aerobic conditions then only 20% was lost in anaerobic conditions. In general, the loss of phosphorus and nitrogen in algal tissues followed the weight loss. As an exception, the amount of nitrogen changed very little during the decomposition of C. glomerata. In field conditions, the photosynthetic activity exceeded the decomposition rate of C. glomerata at lower temperatures in spring. The decomposition of P. littoralis was estimated at 49% of its initial dry weight. The addition of benthic invertebrates had no effect on the decomposition process. In summer, the decomposition rates were estimated at 65% for C. glomerata and 68% for P. littoralis being in the same order of magnitude as observed in laboratory conditions. If the decomposition of C. glomerata was faster at the end of the experiment, the most significant losses of weight of P. littoralis took place during the first 2 weeks of deployment. Idotea baltica significantly contributed to the loss of C. glomerata. The decomposition rate of P. littoralis was reduced by the presence of Mytilus edulis and increased by Gammarus oceanicus.     

The effect of chemotherapy on the in vivo frequency of glycophorin A ‘null’ variant erythrocytes

A human in vivo somatic cell assay based on the enumeration of variant erythrocytes lacking expression of an allelic form of the cell-surface sialoglycoprotein, glycophorin A, was applied to the study of blood samples from patients obtained prior to, during, and following chemotherapy for malignant disease in order to determine the effect of mutagenic chemical agents on the frequency of variant cells. In 22 patients assayed prior to therapy, the mean variant cell frequency was 11.9 per million, which was not significantly different from that observed in healthy controls. In an initial cross-sectional survey, blood samples were obtained at various times during and after therapy from 30 patients diagnosed with a variety of malignancies who were treated with one or more known mutagenic agents including adriamycin, bleomycin, cis-platinum, cyclophosphamide, dacarbazine, etoposide, lomustine, mechlorethamine, melphalan, mitomycin C, and procarbazine. Significant elevations in the mean frequency of variant cells over pre-therapy and normal levels were observed in samples obtained during and after therapy. In a time-series study, 14 breast cancer patients treated with CAF (cyclophosphamide, adriamycin, 5-fluorouracil), CMF (cyclophosphamide, methotrexate, 5-fluorouracil), or VMF (vinblastine, methotrexate, 5-fluorouracil) adjuvant chemotherapy were sampled repeatedly during and after therapy. For the CAF and CMF patients an increase in the frequency of variant cells was observed with a lag in the appearance of induced variants after initiation of therapy; variant frequencies gradually increased during therapy reaching a maximum at or shortly after the end of therapy, then declined to near pre-therapy levels within 6 months. The maximum level of induced variants ranged from 2- to 7-fold over pre-therapy or normal levels depending on the combination of agents used. The breast cancer patients treated with both adriamycin     

The language of systematics,and the philosophy of ‘total evidence’

This contribution analyses the primacy of classification over generalization, and the philosophy of total evidence that emerges from the relation of homology to character statements. Primary conjectures of homology are basic character statements, i.e. statements that predicate properties of organisms, properties that are instantiated by those organisms and/or their parts. Secondary conjectures of homology are embedded in a second‐level (metalinguistic) discourse that turns on the coherence or incoherence of those basic character statements relative to a hierarchy. The coherence or incoherence of character statements is a logical relation between statements, not a causal (historical) relation between organisms. The choice of the hypothesis of relationships that is supported by the largest set of coherent basic character statements is based on the empirical presupposition that the properties referred to by the set of coherent character statements are grounded in causally efficacious relations of inheritance and ontogeny, and co‐instantiated because they are inherited through common ancestry (Hennig's auxiliary principle). Unless that empirical presupposition is causally grounded, phylogeny reconstruction is of an inherently probabilistic nature, whether under parsimony or other models of analysis. The causal grounding of a coherent set of character statements typically seeks a link between character statements and causally efficacious generative mechanisms for morphological characters (as is defeasibly indicated by topology and connectivity), or secondary structure information for molecular characters.     

Histological and cytological study on meristematic nodule induction and shoot organogenesis in Paeonia ostii ‘Feng Dan’

Plant Cell, Tissue and Organ Culture (PCTOC) - This is the first report concerning the sequence of histological and cytological events occurring during organogenesis from cotyledon-derived...     

The play of ‘dirty politics’: ordinary ethics and the evidence of experience on the workfloor in New Delhi,India

This article explores the tensed sociality amongst precariously employed service economy workers in the retail outlets of a prominent eyewear company in New Delhi, India. The bickering staff label the ongoing interpersonal strife ‘dirty politics’. Linked to this are their confident assertions that, in fact, all politics is dirty, and dirty politics is the only type of politics possible – rejecting formal party politics and labour unions as morally vacuous and motivated by utilitarian individualism. Engaging with the anthropology of ordinary ethics, the article demonstrates how the affectively charged experience and talk of petty politicking on the neoliberal workfloor provides critical ‘evidence’ that ‘confirms’ such a politico-ethical view.     

M?ssbauer spectroscopy, electron microscopy and electron diffraction studies of the iron cores in various human and animal haemosiderins

M?ssbauer spectroscopy has indicated significant differences in the iron-containing cores of various haemosiderins. In the present study, haemosiderin was isolated from a number of animal species including man. In addition, haemosiderin was isolated from patients with primary idiopathic haemochromatosis or with secondary (transfusional) iron-overload. The iron cores of the animal and normal human haemosiderin appear to be very similar by M?ssbauer spectroscopy, and the electron diffraction data indicate a ferrihydrite structure similar to that of ferritin cores. The haemosiderin isolated from secondary iron-overload shows anomalous behaviour in its temperature-dependent M?ssbauer spectra. This can be understood in terms of the microcrystalline goethite structure of the cores as indicated by electron diffraction. The haemosiderin cores obtained in the case of primary haemochromatosis have an amorphous Fe(III) oxide structure and show M?ssbauer spectra characteristic of a magnetically disordered material, which only orders at very low temperatures.     

Quantification of the interceptor action of caffeine on the in vitro biological effect of the anti-tumour agent topotecan

Using published in vitro data on the dependence of the percentage of apoptosis induced by the anti-cancer drug topotecan in a leukaemia cell line on the concentration of added caffeine, and a general model of competitive binding in a system containing two aromatic drugs and DNA, it has been shown to be possible to quantify the relative change in the biological effect just using a set of component concentrations and equilibrium constants of the complexation of the drugs. It is also proposed that a general model of competitive binding and parameterization of that model may potentially be applied to any system of DNA-targeting aromatic drugs under in vitro conditions. The main reasons underpinning the proposal are the general feature of the complexation of aromatic drugs with DNA and their interaction in physiological media via hetero-association.     

The effect of chondroitin sulphate–protein on the formation of collagen fibrils in vitro

1. It was found that the precipitation of collagen fibrils at 37 degrees from mixtures of chondroitin sulphate-protein and tropocollagen at physiological ionic strength and pH takes place in two distinct phases. The first occurs immediately on mixing either at 4 degrees or at 37 degrees , and the second occurs only at 37 degrees and after a lag phase whose magnitude depends on the proportions of components. 2. When the second stage of precipitation was inhibited by mixing the reactants at 4 degrees , the initial precipitate was found to contain ;native-type' collagen fibrils and chondroitin sulphate-protein. 3. On the basis of kinetic experiments it was concluded that aggregates of chondroitin sulphate-protein and tropocollagen form instantaneously and that these act as sites for the second stage of precipitation of fibrils. 4. The gels that result after continued incubation at 37 degrees are fibrous in appearance if formed in the presence of the initial precipitate of chondroitin sulphate-protein and tropocollagen. 5. On the basis of these experiments in vitro the authors propose a sequence of events for collagen fibrogenesis in vivo.