Appearance of alpha-smooth muscle actin in human eye lens cells of anterior capsular cataract and in cultured bovine lens-forming cells

Using light and electron-microscopic immunolocalization techniques, and gel electrophoresis combined with immunoblotting, we have examined the expression of cytoskeletal proteins in normal human fetal, child and adult lenses, in human anterior capsular cataract and in bovine lens cells in vivo and in vitro. In this report, we focus our observations on the pattern of actin-isoform expression during normal and pathological situations in vivo and culture conditions. We have noted that cells of developing and mature human lenses as well as bovine lens cells in situ contain only beta- and gamma-actins. In contrast, alpha-smooth muscle (alpha-sm) actin, an isoform typical of smooth muscle differentiation, was demonstrated in bovine lens cells at different times of culture. Moreover, the multilayered cells observed in the subcapsular zone of human anterior capsular cataract were characterized by the presence of alpha-sm actin. Thus, extensive changes in actin-isoform expression take place in lens cells growing in culture and may also occur during cataractogenesis. The biological meaning of the appearance of a marker of myoid differentiation in the ectodermally derived lens-forming cells is discussed.     

Life history of the predatory mite Phytoseius plumifer and the effect of nutrition on its biology (Acarina: Phytoseiidae)

Phytoseius plumifer could develop and reproduce when fed on the different stages of the red spider mite T. cinnabarinus. The female immature stages lasted for an average of 4.8, 3.8 and 4.5 days respectively when fed on eggs, immatures and adults of the prey at an average temperature of 29.5±1°C. The average number of deposited eggs per female was significantly greater when it fed on immatures and adults (about 45 eggs) than when it fed on prey eggs (about 29 eggs). During the whole life-span the predator female fed on an average of 969 eggs or 438 immatures or 346 adults, of these more than 93% were consumed during the adult stage. Males lived for shorter time and consumed fewer prey than females.The predator could develop and reproduce successfully on date-palm pollen but at a slower rate. On this diet the immature stages lasted 10.5 days and female laid an average of 16.6 eggs. On hollyhock and cotton pollen, or sweet potato leaves, larvae failed to develop to adults.

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Zusammenfassung Phytoseius plumifer konnte sich entwickeln und fortpflanzen, wenn er mit verschiedenen Stadien der Roten Spinnmilbe, Tetranychus cinnabarinus, gefüttert wurde. Die Entwicklungs-stadien des Weibchens dauerten bei einer Mitteltemperatur von 29,5±1° je nach Fütterung mit Eiern, Jungtieren oder Adulten der Beute durchschnittlich 4,8, 3,8 oder 4,5 Tage. Die durchschnittliche Anzahl von einem Weibchen abgelegter Eier war signifikant größer, wenn sie mit Larven und Erwachsenen gefüttert wurden (etwa 45 Eier), als wenn sie mit Eiern der Beutetiere ernährt wurden (etwa 29 Eier). Während der Gesamtlebensdauer verzehrte ein Raubmilben-Weibchen im Durchschnitt 969 Eier oder 438 unreife oder 346 erwachsene Beutetiere, von denen mehr als 93% während des Imaginalstadiums verbraucht wurden. Männchen lebten kürzere Zeit und vertilgten weniger Beute als die Weibchen.Der Räuber konnte sich an Dattelpalmen-Pollen entwickeln und erfolgreich fortpflanzen, jedoch mit einer geringeren Rate. Bei dieser Ernährung dauerte die Entwicklung bis zur Imago 10,5 Tage und ein Weibchen legte im Durchschnitt 16,6 Eier. Mit Stechpalmen- und Baumwoll-Pollen oder an Süßkartoffel-Blättern entwickelten sich die Larven nicht bis zur Imago.

     

Human arterial smooth muscle cells in culture: Inverse relationship between proliferation and expression of contractile proteins

Summary Human arterial smooth muscle cells (hASMC) from explants of the inner media of uterine arteries were studied in secondary culture. We had previously found that these cells depend on exogenous platelet-derived growth factor (PDGF) for proliferation in vitro. Deprivation of the serum mitogen(s) by culture in plasma-derived serum or bovine serum albumin (BSA) caused a true growth arrest that was reversible upon reexposure to the mitogen(s). When added to serum-containing medium, heparin caused a reversible growth arrest which could be competed for by increasing concentrations of serum. In the current study we used a set of smooth muscle-specific actin and myosin, antibodies to study the expression of contractile proteins in stress fibers under indirect immunofluorescence on hASMC in culture. Even in sparse culture, grwoth-arrested hASMC expressed stress fibers containing these actin and myosin epitopes. This was true irrespective of whether growth arrest was achieved by culture in media containing only BSA or a combination of heparin and whole blood serum. hASMC proliferating in whole blood serum in sparse culture did not express such strees fibers, as judged by immunofluorescent staining. This was true also for cells that were restimulated to proliferate in serum after a growth arrest. Utilizing a monoclonal antibody against a nuclear antigen expressed in proliferating human cells, we were able to demonstrate an inverse relationship between the expression of this antigen and the SMC-specific contractile proteins, respectively. Under these culture conditions, the reversible transition between defifferentiated and differentiated hASMC was almost complete and terminated about 1 wk after the change in culture condition. We conclude that hASMC in vitro respond, to exogenous PDGF by proliferation and dedifferetiation as a single population of cells. We also conclude that this modulation is reversible, because the cells become uniformly quiescent and differentiated when the mitogenic stimulus is blocked or removed. This study was supported by grants from the Swedish Medical Research Council (Project no. 4531 and 6816), the Swedish Association against Heart and Chest Diseases, the King Gustaf V and Queen Victoria Foundation, the National Institutes of Health, Bethesda, MD (grant HL 29873) and the Swedish National Board for Laboratory Animals.     

Alpha-smooth muscle actin is expressed in a subset of bone marrow stromal cells in normal and pathological conditions

A series of 217 trephine bone marrow biopsies from adult patients and specimens from 16 fetuses and 5 infants were examined for the presence of stromal myoid cells (MCs) using a monoclonal antibody recognizing alpha-smooth muscle actin. In the normal adult bone marrow, stromal cells did not contain alpha-smooth muscle actin, whereas during fetal life, many alpha-smooth muscle actin-containing MCs were connected with vascular sinusoids in the primitive bone marrow. This cell type reappeared in various characteristic distribution patterns in adult bone marrow during different neoplastic and non-neoplastic conditions including metastatic carcinoma, Hodgkin's disease, multiple myeloma, hairy cell leukemia, acute myeloid leukemia (FAB M4, 5, 7) and chronic myelo-proliferative diseases. In general, the appearance of MCs was associated with a slight to pronounced increase in the deposition of reticulin and collagen fibers. We propose that bone marrow MCs represent a distinct subpopulation of fiber-associated or adventitial reticular cells undergoing cytoskeletal remodeling in response to various stimuli.     

Three-dimensional geometrical and mechanical modelling of the lumbar spine.

The main objective of this study is to design a three-dimensional geometrical and mechanical finite element model of the lumbar spine. The model's geometry is constructed using six parameters per vertebra. These parameters are digitized from two X-rays (anterio-posterior and lateral), thus yielding an individualized model which can be arrived at from the radiographs of a tested specimen. This procedure makes the model validation easier, as geometry is generally a factor of dispersion in experimental results. The geometrical reconstruction, in the form of a finite elements mesh, was effected for the whole lumbar spine. The global coherence of the model was verified.     

Action of general and alpha-smooth muscle-specific actin antibody microinjection on stress fibers of cultured smooth muscle cells

Arterial smooth muscle cells express alpha- and gamma-smooth muscle, as well as beta- and gamma-cytoplasmic actins. Two actin antibodies, one recognizing smooth muscle and cytoplasmic actin isoforms, the other recognizing specifically alpha-smooth muscle actin, were microinjected into cultured aortic smooth muscle cells. The effect of these antibodies on stress fiber organization was examined by staining with rhodamine-labeled phalloidin and by immunofluorescence with the same antibodies. Microinjection of the general actin antibody abolished most of the stress fiber staining with all reagents, but did not significantly affect the shape of the injected cells. This suggests that stress fiber integrity is not absolutely necessary for the maintenance of cell shape within the time of observation. Microinjection of the specific alpha-smooth muscle antibody abolished to various extents the staining of stress fibers with this antibody, but left practically intact their staining with rhodamine-labeled phalloidin and with the general actin antibody. This suggests that the incorporation of alpha-smooth muscle actin is not absolutely necessary for the maintenance of stress fiber integrity in cultured smooth muscle cells.     

Evidence for distinct polygenic regulation of antibody responses to some unrelated antigens in lines of mice selected for high or low antibody responses to somatic antigen of Salmonella

The effect of the selective breeding of mice for high or low antibody production to complex immunogens is largely nonspecific, since it modifies the responsiveness of high (H) and low (L) lines to many antigens unrelated to the selection antigen. However, the nonspecific effect of the polygenic control operating in these lines is not a general feature. For example, the group of genes in selection IV, carried out for responsiveness to somatic antigen of Salmonella, does not modify the responses to sheep erythrocytes (SE). Despite equivalent responses in H and L mice of selection IV, a large variability was found in individual responses of F₂ interline hybrids, which demonstrates the presence of alleles with high or low effect on responses to SE. A selective breeding (Selection IV-A) was therefore initiated from this F₂ population for responsiveness to SE. A progressive interline divergence occurred during the first seven generations of selection; the interline separation was due to polygenic regulation (about four independent loci from a preliminary estimate).Equivalent responses to the s antigen of Salmonella are observed in the two lines. This constitutes additional evidence for distinct polygenic regulation of responses to SE and to somatic antigen. Moreover, the pattern of responses to several unrelated antigens (nonspecific effect) also differs between Selections IV and IV-A.Abbreviations H high responder lines - L low responder lines - s somatic antigen of Salmonella - f flagellar antigen of Salmonella - R response to selection - S selection differential - F₀ foundation population - h² heritability (realized) - RGG rabbit gamma globulin - CE chicken erythrocyte - HE human erythrocyte - PE pigeon erythrocyte - SE sheep erythrocyte     

An anatomical subject-specific FE-model for hip fracture load prediction

In order to reduce the socio-economic burden induced by osteoporotic hip fractures, finite element models have been evaluated as an additional diagnostic tool for fracture prediction. For a future clinical application, the challenge is to reach the best compromise between model relevance and computing time. Based on this consideration, the current study focused on the development and validation of a subject-specific FE-model using an original parameterised generic model and a specific personalization method. A total of 39 human femurs were tested to failure under a quasi-static compression in stance configuration. The corresponding FE-models were generated and for each specimen the numerical fracture load (F _FEM) was compared with the experimental value (F _EXP), resulting in a significant correlation (F _EXP = 1.006 F _FEM with r ² = 0.87 and SEE = 1220 N, p < 0.05) obtained with a reasonable computing time (30 mn). Further in vivo study should confirm the ability of this FE-model to improve the fracture risk prediction.