Evidence that sister chromatid exchanges and chromatid breaks are two independent events

The relative frequencies of sister chromatid exchanges (SCE) and chromatid breaks in BrdU (5-bromodeoxyuridine) — sensitive site (lq22 lq23) in Chinese hamster cells after BrdU incorporation were studied. The results show that chromatid breaks do not follow the exchange hypothesis and provide evidence that chromatid breaks and SCEs are two independent events despite some common features.     

Sister chromatid exchanges are preferentially induced at expressed and nonexpressed common fragile sites

Summary To investigate the relationship between common fragile sites and sister chromatid exchange (SCE), lymphocyte cultures were treated with aphidicolin and bromodeoxyuridine (BrdU) and analyzed using a sequential GSCE staining protocol. A total of 1163 SCEs were mapped to their corresponding G-band sites, which were assigned to one of the following four categories: fragile sites expressed; fragile sites nonexpressed; nonfragile sites with breaks; or nonfragile sites with no breaks. The designated common fragile sites were found to be preferred locations for SCE formation, not only when these sites were expressed as visible gaps or breaks, but even when they were nonexpressed in the cell. SCEs were also more likely to occur at nonfragile sites with breaks than at nonfragile with no break sites. Further, SCEs were found to be distributed nonrandomly across fragile sites and nonfragile sites, and among the fragile sites, the high frequency SCE sites were highly correlated with the high frequency breakage sites. These data support the hypothesis of common steps in the mechanism of aphidicolin-induced SCE formation and common fragile site expression.     

Evidence for chromosomal replicons as units of sister chromatid exchanges

Chromosomal replicons have been described as the cytological counterpart of DNA replicon clusters and have previously been studied in vitro using premature chromosome condensation-sister chromatid differentiation (PCC-SCD) techniques. Chromosomal replicons are visualized as small SCD segments in S-phase cells, and measurement of these segments can provide estimates of relative chromosomal replicon size corresponding to DNA replicon clusters functioning coordinately in S-phase. Current hypotheses of sister chromatid exchange (SCE) formation postulate that sites of SCE induction are associated with active replicons or replicon clusters. We have applied the PCC-SCD technique to in vivo studies of mouse bone marrow cells that have been treated with cyclophosphamide (CP) for two cell cycles. We have been able to visualize chromosomal replicons, as well as SCEs which have been induced in vivo by CP treatment, simultaneously in the same cells. Chromosomal replicons visualized as small SCD segments were measured in PCC cells classified at early or late S-phase based on SCD segment size prevalence. Early S-phase (E/S) PCC cells contained 90% of the SCD segments measured clustered in a segment size range of 0.1 to 0.8 m with a peak value around 0.3 to 0.6 m regardless of CP treatment. As the cells progressed through S-phase, late S-phase (L/S) PCC cells were characterized by the appearance of larger SCD segments and even whole SCD chromosomes in addition to small SCD segments. A concentration of units around 0.4 to 1.0 m was found for L/S SCD segment size distributions regardless of CP treatment with an apparent bimodal profile. Our in vivo data support the existence of a subunit organization of chromosomal replication with a basic functional unit being 0.3 to 0.6 m in size. In addition, we have found that this chromosomal unit of replication or chromosomal replicon does not seem to be functionally perturbed by the mutagen CP. We also found that small SCD segments of 0.4 to 0.7 m in length were involved in the formation of an SCE, suggesting that both spontaneous and CP-induced SCEs occur between chromosomal replicons. These findings provide direct cytogenetic evidence to support a replicon cluster/chromosomal replicon model for SCE formation.     

Die Intravitale Karyorrhexis Der Exokrinen Pankreaszelle im Elektronenmikroskopischen Bild

Zusammenfassung Die intravitale Karyorrhexig wird am exokrinen Pankreas der Maus nach Osmium- und Formalinfixierung licht- und elektronenmikroskopisch untersucht. Im Verlaufe der Karyorrhexis kommt es zu einer Trennung des Karyoplasmas in 'Chromatin und 'Karyolymphe (Interchromatin-Substanz). In fortgeschrittenen Stadien ist das Bild solcher Zellkerne — im Gegensatz zu normalen Intermitosekernen — nach beiden Fixierungsmethoden fast identisch. Das Chromatin ist in dichten, membranständigen Kappen konzentriert. Der übrige Kernraum wird — abgesehen vom Nukleolus — von der hellen Karyolymphe ausgefüllt. In ihrem Bereich treten die sog. Interchromatingranula (200–250 Å), einzeln oder in dichten Haufen, meist deutlich hervor. Der Nukleolus, dessen Feinstruktur nicht auffallend verändert ist, liegt entweder frei oder den Chromatinkappen unmittelbar an. Die Kernhülle ist bei fortgeschrittener Karyoplasmaentmischung unversehrt oder über kürzere oder längere Strecken geschwunden. Im Endstadium der Karyorrhexis sind mehrere, von Kernmembranfragmenten unvollständig begrenzte Chromatinbrocken wahllos im Zelleib verstreut. Die cytoplasmatischen Strukturen können auch bei ausgeprägten Kernveränderungen noch weitgehend normal aussehen.Die karyorrhektischen Kernveränderungen werden als Folge einer 'Entmischung des Karyoplasmas gedeutet, für die eine Dehydratation der DNS-tragenden Chromosomenstrukturen verantwortlich gemacht wird. Auch das Sichtbarwerden und das Verklumpen der 'Interchromatingranula wird auf eine Dehydratation zurückgeführt.

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Summary The intravital caryorrhexis in the exocrine pancreas of the mouse was studied by means of light- and electron microscopy after fixation with osmium and formaldehyde. During caryorrhexis the caryoplasm segregates into chromatin and caryolymphe (interchromatinic substance). In contrast to normal intermitotic nuclei both fixations result in a very similar picture of the typical caryorrhectic nuclei. The chromatin is concentrated in dense caps associated with the nuclear envelope. Except those dense caps and apart from the nucleolus the whole nucleus is occupied by the light caryolymphe. In this area the so-called interchromatinic granules (200–250 Å) become clearly visible, isolated or in dense clumps. The nucleolus, the fine structure of which has not remarkably changed, can be found either isolated or in touch with the chromatin caps. During advanced segregation of the caryoplasm the nuclear envelope was found to be either unimpaired or had disappeared for variable lenghths. In the final stage of caryorrhexis several chromatin clumps — partly confined by fragments of the nuclear envelope — are distributed irregularly within the cell. Even in the case of pronounced nuclear changes the cytoplasmic structures may appear almost unaffected.It is assumed that the nuclear changes during caryorrhexis are due to a segregation of the caryoplasm, caused by dehydration of the DNA-carrying chromosome fibrils. Also dehydration is made responsible for the appearance and clumping of the interchromatinic granules.

     

On the heterogeneity of the slow reassociating (“unique”) DNA

Summary The slow reassociating fraction of mouse DNA (unique DNA), when allowed to reassociate in 0.14 m sodium phosphate buffer at 50 °C showed a biphasic melting curve with a transition at 78–80 °C. On the basis of this feature, the slow reassociating DNA was separated preparatively into two fractions: unique DNA I and II. Their duplexes showed differences with respect to thermal stability, S1 nuclease resistance and rate of reassociation. About one third of the sequences in each fraction were fraction-specific. The conclusion was drawn that for unique DNA I these should be the low repetitive or single copy related sequences (multigene families) and for unique DNA II—the unrelated single copy sequences or recent families of low repetitive not yet diverged sequences.     

Effect of androgens on histochemical fibre type

Summary The temporal muscles of the guinea pig show a sexual differentiation reflected in their histochemical enzyme pattern. Using histochemical methods for mitochondrial (SDH, -GPDH), and glycolytic enzymes (phosphorylase, LDH) it could be shown, that in adult animals the male muscle is a white muscle with marked activity of glycolytic enzymes, the female muscle a red muscle displaying high activity of mitochondrial enzymes. This differential enzyme pattern can be converted by the application of testosterone to the female type during the postnatal development. The male sex hormone thus affects the histochemical enzyme pattern of the muscle, converting the red, female into a white, male muscle in the female guinea pig.     

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.     

Contextualizing Images in Dreams and Daydreams

A contextualizing image (CI) is a powerful central image of a dream which appears to contextualize (provide a picture-context for) the dreamer's emotion. For instance, dreamers who have experienced any serious traumatic event sometimes dream, I was overwhelmed by a tidal wave. This appears to picture their feeling of terror and/or vulnerability.A scoring system for CIs is examined here and is applied to dreams and daydreams supplied by 40 students. Two raters scoring dreams on a blind basis showed good inter-rater reliability. Recent dreams were shown to have more as well as more intense CIs than recent daydreams; likewise, dreams that stand out had more intense CIs than daydreams that stand out. Students with thin boundaries had more and more intense CIs than students with thick boundaries in their recent dreams and nightmare, but not so clearly in dreams and nightmares that stand out. The emotions judged as contextualized by the powerful images tended towards fear/terror and helplessness/vulnerability in dreams (especially in dreams that stand out) whereas emotions contextualized by images in daydreams showed a wide range with no clusters.     

Comparative residual toxicities of pesticides to the predator Agistemus industani (Acari: Stigmaeidae) on citrus in Florida

Residual toxicities of registered and selected experimental pesticides used on citrus against Agistemus industani Gonzalez (Acari: Stigmaeidae) were compared. Pesticides considered highly toxic to A. industani were: abamectin 0.15 EC at 731ml/ha+FC 435-66 petroleum oil at 46.8l/ha, pyridaben 75WP at 469g/ha, ethion 4EC at 7.01l/ha+FC 435-66 petroleum oil at 46.8l/ha, propargite 6.55 EC at 3.51l/ha, chlorfenapyr 2SC at 1.46l/ha applied alone or in combination with FC 435-66 petroleum oil at 46.8l/ha, sulphur 80DF at 16.81kg/ha, dicofol 4EC at 7.01l/ha, fenbutatin oxide 50WP at 2.24kg/ha, benomyl 50WP at 2.24kg/ha, benomyl 50WP at 1.68kg/ha+ferbam 76 GF at 5.60kg/ha, ferbam 76GF at 11.21kg/ha, neem oil 90EC at 46.8l/ha, and copper hydroxide DF (40% metallic copper) at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha. Pesticides that were moderately to slightly toxic included: copper sulphate 98% at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha, fenbuconazole 2F at 280ml/ha+FC 435-66 petroleum oil at 46.8l/ha, FC 435-66 petroleum oil applied alone at 46.8l/ha or 23.4l/ha, and diflubenzuron 25WP at 1.40kg/ha. Pesticides that were non-toxic included: fenbuconazole 2F at 585ml/ha, malathion 57EC at 5.85l/ha, FC 435-66 petroleum oil at 46.8l/ha, carbaryl 80S at 3.36kg/ha, chlorpyrifos 4EC at 4.68l/ha, and formetanate 92SP at 1.12kg/ha. Understanding the toxic effects of field weathered pesticides against key predacious mite species is important for effective IPM. The results of this study provide a comparison of direct and indirect toxic effects of various pesticides to A. industani under field conditions.     

Male swarms at landmarks and scramble competition polygyny inPolistes gallicus (Hymenoptera: Vespidae)

At the end of summer, males of Polistes gallicusfly in swarms around vertical landmarks and land in clusters on their favorite perches, where they drag their legs and abdomen. Here males occasionally crowd around a perched female; they make no effort to defend an exclusive mating territory but instead attempt to copulate by displacing rivals from the female. In this work we describe this spatial-nuptial system, which entails site fidelity without territoriality, unisexual swarms, common patrol routes, collective sexual approaches, and scramble competition polygyny. Mating success is evaluated in relation to the familiarity with flight paths (routine patrollers versus newcomers), to the type of sexual approach (single males versus in- group males), and, in the laboratory, to the individual activity level.     

Electron microscopic and cytochemical study on the role of Golgi elements and plasma membrane of enterocytes in the intestinal lipid transport

Summary Intestinal lipid absorption and transport were investigated in albino rats. The observations point towards the existence of a continuity between plasma membrane invaginations and elements of the Golgi complex on its mature face. They also suggest a segregation of lipid droplets by paired Golgi membranes and plasma membrane invaginations. The following way for lipid transport is deduced: lipid droplets moving inside the smooth endoplasmic reticulum accumulate progressively and are condensed in Golgi cisternae of the forming face. Their limiting membrane ruptures and liberated lipid droplets are segregated by paired Golgi membranes of the mature face or by plasma membrane invaginations. Subsequently the inner of the two segregating membranes disappears while the lipid droplet is moved towards the intercellular space inside a canal communicating with this space. The suggestion is made that the Golgi apparatus is of double origin: one component representing a terminal plication of the endoplasmic reticulum; the second one—a terminal plication of the plasma membrane invagination. This concept explains the ultrastructural and histochemical differences between Golgi membranes of the forming and mature faces of the complex.     

Age-related accumulation of lysosomes and other cytological features in active thyroid follicles of the CBA mouse

Summary This study attempts to elucidate the mechanism through which lysosomal accumulation occurs with age in the epithelial cells of the thyroid gland and especially in the active follicles of the aging mouse thyroid. Thyroid morphology and function in old CBA (at least 24 months of age) male mice were compared with those in young (2 months of age) animals. The effects of different intake of iodine were tested and compared in both cohorts, each of which was divided into three groups: (i) low iodine group, (ii) moderate iodine group, and (iii) high iodine group. As expected, the present work confirmed the well-known accumulation with age of cold follicles coexisting with active follicles in the old mouse thyroid. Attention has been focused on the active follicles whose follicular cells contained in their cytoplasm a large number of pleomorphic dense bodies. The lysosomal nature of these bodies, referred to as secondary lysosomes, was confirmed by histochemistry; however, they displayed variability in acid phosphatase staining. In old animals, regardless of the type of iodine regimen, the ratio between relative follicular volume and relative colloid volume as determined by morphometry remained unchanged. Ultrastructurally, the relative volume occupied by secondary lysosomes in active follicles was always higher than in the young groups. Autoradiographic studies with ¹²⁵I revealed that a large part of the radioactivity was located in secondary lysosomes of thyroid cells in active follicels of old mice when radioiodine was injected 3 weeks before death. Two different types of vacuoles were present in a non-negligible number of thyrocytes of the active follicles in aged cohorts. The first type was made up of grossly dilated rough endoplasmic cisternae, the second corresponded to intracytoplasmic microfollicular vacuoles. Both aspects have been described in conditions of chronic stimulation. It is concluded (1) that different intake of iodine for 6 weeks does not modulate the thyroid morphology in old mice; (2) that in the thyrocytes of the active follicles in old mice accumulation of secondary lysosomes occurs due to a slowdown of turnover; and (3) that the follicular cells of active follicles feature morphological aspects suggesting a hyperactive state compensating the lack of hormone production in the cold follicles.     

Polymorphism of the HLA DR1 haplotype in the Israeli population investigated at the serological,cellular, and genomic levels

In the present report, we used serological, cellular, and restriction fragment length polymorphism (RFLP) to investigate the DR1 haplotype in the Israeli population. We describe an Israeli homozygous typing cell (HTC), HLA-DwLVA, which defines a new lymphocyte-activating determinant associated with Bw65, DR1 and distinct from Dwl. The parents of this donor, non-Ashkenazi Algerian Jews, are first cousins and share HLA-Cw8, Bw65, BfS, DR1, DQw1, DPw4. No specificity could be assigned to HLA-DwLVA using the 91 Ninth Workshop HTCs. Two families and forty unrelated DR1 individuals were studied with DwLVA and a panel of DR1/Dw1 HTCs. HLA-DwLVA showed segregation as a single determinant within families. This new specificity was present in 24 out of 40 (60%) unrelated DR1 individuals, indicating that in the Israeli population DwLVA is the main lymphocyte-defined determinant associated with the serologically defined DRI specificity, in contrast to non-Jewish Caucasoids where DR1 is significantly associated with Dw1. The vast majority of DwLVA-positive carriers were also Bw65 carriers, indicating that Bw65, DR1, DwLVA may represent a typical allele combination in the Israeli population. The RFLP analysis established the correlation of certain RFLPs with Dw1 and DwLVA. In addition, we describe a cluster of RFLPs that may correspond to a new Dw subtype associated with DR1, for which no serological and cellular reagents have been described so far.     

The effect of intracapillary media feed protocols on hollow fiber cell culture

Summary Two intracapillary (IC) media feed protocols termed media rich and media lean were examined in an effort to understand the effect of this variable on hollow fiber cell cultures. The media rich protocol emphasized a high volume IC media per day (5 liters) containing no serum and a normal amount of extracapillary (EC) media serum (10% v/v). Alternatively, the media lean protocol used up to 1.0 liter of IC media per day containing 5% v/v serum and increased EC media serum (20% v/v). Both protocols produced substantial amounts of antibody in 25 days using HFN7.1 hybridoma cells (ATCC CRL 1606), however the media rich protocol produced twice as much antibody as the media lean protocol. The metabolism of the cells was dramatically different as measured by glucose uptake rate (GUR) with media lean cells having a six-fold lower GUR. Our results indicate that the media rich protocol is useful for producing larger amounts of antibody in a short time frame. The media lean protocol may be considered when the production costs of antibody, particularly media and serum, is the overriding concern.     

Short- and long-term effects of ACTH on the adrenal zona glomerulosa of the rat

Summary Short-term ACTH treatment provoked a decrease in volume of the lipid-droplet compartment in rat zona glomerulosa cells, and a rise in plasma and intracellular concentrations of corticosterone and aldosterone. It enhanced activities of 3-hydroxysteroid dehydrogenase (3HSD), 11-hydroxylase (11OH) and 18-hydroxylase (18OH). Long-term ACTH administration produced a hypertrophy of the zona glomerulosa and its parenchymal cells, a result of the increase in volume of the smooth endoplasmic reticulum and the mitochondrial compartment. The surface area per cell of mitochondrial inner membranes increased; the tubular cristae were transformed into a homogeneous population of vesicles. The plasma and intracellular concentrations of corticosterone further increased, whereas those of aldosterone fell below basal levels (the aldosterone-escape phenomenon). The activities of 3HSD and 11OH were enhanced, that of 180H decreased. Therefore, ACTH stimulates zona glomerulosa growth and transforms parenchymal elements into zona fasciculata celltypes. Cyanoketone nullified acute ACTH effects on plasma and intracellular concentrations of corticosterone and aldosterone, but did not affect the activities of 11OH and 18OH. Chronic ACTH treatment produced similar results, although 18OH activity was not suppressed. The mechanism underlying the aldosterone-escape phenomenon may thus involve a rise in the intracellular concentration of corticosterone, caused by the enhanced synthesis and activation of 3HSD and 11OH.     

Assignment of human ferritin genes to chromosomes 11 and 19q13.3→19qter

Summary Extracts of hamster-human and mouse-human hybrids, some with translocations involving chromosome 19, have been assayed for both human spleen ferritin (rich in L subunits) and human heart ferritin (rich in H subunits). Hybrid lines retaining part of the long arm of chromosome 19 including the region 19q13.319qter produced human L type ferritin. This confirms the previous assignment of the ferritin gene to chromosome 19 (Caskey et al. 1983). However, lines retaining chromosome 11 were found to contain human H type ferritin suggesting that the gene for the H subunit is on this chromosome. The presence of chromosome 6 was not necessary for the expression of either H or L type human ferritin. It thus seems unlikely that the gene for idiopathic haemochromatosis is a ferritin gene.     

Line source models for active fibers

It is shown that the utility of usual circular line source models is highly questionable. A line source model related to arbitrary cross-sections is given and essential differences between the source strength of the circular and non-circular model are found in the case of fairly flat elliptic fibers.     

We Do Not Dream of the 3 R's: Implications for the Nature of Dreaming Mentation

This report examines the extent to which dream recall involves the 3 R's (reading, writing, and arithmetic). Two separate studies were done. In the first study, two scorers rated, on a blind basis, a total of 456 written dream reports, available from five previous studies. There was perfect agreement between the two scorers. They agreed that there were no instances of reading, no instances of writing, and one instance of probable calculating in the 456 dreams. The second study was a questionnaire survey. Complete responses were obtained from 240 frequent dreamers (who reported remembering a mean of seven dreams per week). The study examined in two ways the frequency of the 3 R's in their recalled dreams. First, in answer to direct questions as to how frequently they dreamt about each activity, roughly 90% of the respondents reported that they never or hardly ever dreamt about each of four activities: reading, writing, typing, and calculating. In answers to other questions, this group reported spending a mean of six hours per day engaged in these activities. Second, responses as to the relative prominence of six activities (walking, writing, talking with friends, reading, sexual activity, typing) in dreaming versus waking produced two clear groupings of activities. Walking, talking with friends, and sexual activity were each rated almost as prominent in dreaming as in waking whereas the second group consisting of writing, reading, and typing were rated as far more prominent in waking than in dreaming. The two activity groups differed at p < .0001. Thus, the 3 R's appear to occur very infrequently in dreams. These findings are placed in a theoretical frame which suggests that dreaming (compared to waking) deals very little with serial activities characterized by input—rapid-processing—output in which the neural nets function in a feed-forward mode. Rather, dreaming may be characterized by relatively broad or loose connection making in which the nets function more in an autoassociative mode.     

Multiple rough endoplasmic cisternae in “C” cells

Summary Multiple rough endoplasmic cisternae were found in C cells of the adult rat, at interphase. They are considered to be normal constituents of C cells. Their morphological relation to rough endoplasmic reticulum and their close proximity to mitochondria, Golgi dictyosomes and secretory granules suggest that they may have a role in the secretory activity of this endocrine cell.