Molecular mechanisms in the control of limb regeneration: the role of homeobox genes

The Ca2+ sensitivity of cardiac myofibrillar force production can be decreased by acidosis or inorganic phosphate (P(i)) and increased by caffeine. To investigate whether the source of tissue influences the potency of these agents, we compared the actions of acidosis (change of pH from 7.0 to 6.2), P(i) and caffeine (both 20 mM) on force production of skinned cardiac muscles from adult ventricle, adult atrium and neonate ventricle of the rat. Maximum Ca(2+)-activated force was reduced by all three interventions and the responses of the different muscle types to a given intervention were similar. Acidosis reduced myofibrillar Ca2+ sensitivity by 1.09 and 1.04 pCa units in adult ventricle and atrium, respectively, and P(i) reduced it by 0.19 and 0.22 pCa units. However, each effect was only one-third as great in the neonate ventricle, which showed falls of 0.33 pCa units for acidosis and 0.06 for P(i). In contrast, caffeine raised the Ca2+ sensitivity by the same amount (approximately 0.4 pCa units) in all three muscle types. The differential effect between adult and neonate seen with both acidosis and P(i) suggests some similarity in the mechanisms by which these factors decrease Ca2+ sensitivity. In contrast, the equal effects of caffeine on neonate and adult suggests that caffeine acts by a completely different mechanism. The lower pH- and P(i)-sensitivity of the neonatal ventricle can help to explain why neonatal and adult myocardium exhibit differential force responses to ischaemia (or hypoxia alone).     

Symbioses in Amphipholis squamata (Echinodermata, Ophiuroidea, Amphiuridae): geographical variation of infestation and effect of symbionts on the host's light production

Populations of the polychromatic and bioluminescent species Amphipholis squamata from eight locations were examined for internal and external symbionts. At three locations (two in the United Kingdom and one in Papua New Guinea), no symbionts were present, while four species were recovered from the remaining locations: Cancerilla tubulata and Parachordeumium amphiurae (copepods), Rhopalura ophiocomae (orthonectid) and an undescribed species of rhabdocoel turbellarian. No ophiuroid individual hosted more than one symbiont species, despite the presence of two or more within a population. Symbiont presence and prevalence varied with location, and with colour variety, but with no apparent pattern or trends. Light-production characteristics of the host were affected by the presence of all symbionts except C. tubulata. These effects, however, did not vary between colour varieties or between geographical locations, but were specific to the symbiont species: the presence of P. amphiurae resulted in enhanced intensity of light production, while that of R. ophiocomae and the turbellarian species resulted in reduced intensity. The kinetics of light production (time until maximum output) were altered only by the presence of the turbellarian. Changes in the light-production characteristics are discussed in relation to morphological, energetical and physiological effects of the symbioses.     

The fine structure of the embryo during the gastrula stage of Comanthus japonica (Echinodermata: Crinoidea)

The fine structure of the embryo of Comanthus has been described by scanning and transmission electron microscopy at two-hourly intervals throughout the gastrula stage (from the fifth through the fifteenth hours of development). At 5 hr, gastrulation has occurred in the absence of any structure comparable to the echinoid hyaline layer; therefore, at least one important mechanism proposed for echinoid gastrulation cannot occur in this crinoid. At 7 hr, the blastocoelic basal lamina has formed, and all ectodermal and entodermal nuclei contain dense aggregates, which are probably perichromatin fibrils. At 9 hr, the blastocoel contains mesenchyme cells, presumably of entodermal origin. At 11 hr, ciliogenesis has started at the apical surfaces of the ectoderm cells and at the archenteral surfaces of the entoderm cells; many of the newly formed cilia are swollen subterminally. At 13 hr, a conspicuous glycocalyx is beginning to cover the apical ends of the ectoderm cells, and the fertilization membrane is beginning to dissolve from its inner surface. Between 5 and 13 hr, there is a gradual development of a junctional complex associating the apicolateral margins of the ectoderm cells; the zonula adherens part of the complex appears at 5 hr and is well developed by 7 hr, and then the septate junction part of the complex appears at 9 hr and is well developed by 13 hr. At 15 hr, the blastopore has closed, the ectodermal glycocalyx is fully developed, some mesenchyme cells appear to be differentiating into skeleton forming cells, and the fertilization membrane is in the last stages of dissolution.     

Ontogeny of the endocrine cells of the intestine and rectum of sea bass (Dicentrarchus labrax L.): an ultrastructural study

Several endocrine cell types were ultrastructurally characterized during the differentiation of the intestine and rectum of sea bass (Dicentrarchus labrax L.) larvae. Only one cell type (type I) was found in the posterior region of the undifferentiated gut of 5-day-old larvae (phase I). Types V and VI were found in both the intestine and rectum, types II, III and IV in the intestine, and types VII and VIII in the rectum of 9- and 12-day-old larvae (phase II), the rectum alone showing signs of functional differentiation. In phase III larvae, in which both the intestine and rectum were differentiated, types IX, X, XI, XII, XIII, XIV and XV were found in the intestine, only types X, XI and XII being seen in the rectum. Besides these, a new cell type, XVI, was observed in the intestine of 55- and 60-day-old larvae (phase IV), in which the digestive tract was completely differentiated. The endocrine cells appearing in phases I and II showed very scarce secretory granules and the ultrastructural features of undifferentiated cells. Some endocrine cell types in the earliest developmental stages were related to some of those found later. A maturational process of the endocrine cell types paralleled the differentiation of the intestine and rectum, with an apparent increase in the number of secretory granules accompanying organelle development.     

Cellular mechanisms for developmental toxicity of chlorpyrifos: targeting the adenylyl cyclase signaling cascade

Developmental neurotoxicity caused by chlorpyrifos exposure is generally thought to target cholinesterase but chlorpyrifos may also act on cellular intermediates, such as adenylyl cyclase, that serve global functions in the coordination of cell development. In the current study, neonatal rats were exposed to apparently subtoxic doses of chlorpyrifos (no weight loss, no mortality) either on Postnatal Days 1-4 or on Postnatal Days 11-14, and the effects on components of the adenylyl cyclase cascade were evaluated in brain regions that are enriched (forebrain) or sparse (cerebellum) in cholinergic innervation, as well as in a nonneural tissue (heart). In all three, chlorpyrifos evoked deficits in multiple components of the adenylyl cyclase cascade: expression and activity of adenylyl cyclase itself, functioning of G-proteins that link neurotransmitter and hormone receptors to cyclase activity, and expression of neurotransmitter receptors that act through this cascade. Disruption of signaling function was not restricted to transduction of cholinergic signals but rather extended to adrenergic signals as well. In most cases, the adverse effects were not evident during the immediate period of chlorpyrifos administration, but appeared after a delay of several days. These results suggest that chlorpyrifos can affect cell development by altering the activity and reactivity of the adenylyl cyclase signaling cascade, a major control point for trophic regulation of cell differentiation. The effects are not restricted to cholinergic targets, nor even to the central nervous system. Hence, disruption of cell development by chlorpyrifos is likely to be more widespread than previously thought.     

X-ray diagnosis of Crohn's disease in the intestine (author's transl)

The article describes the essential roentgenologic findings associated with Crohn's disease in the intestine, manifested as stenoses, thickening of the walls, fistula formation and pseudodiverticula or appearance of Bodart's intermediary segment, on the basis of characteristic findings, followed by valuation in respect of the exclusive possibility of diagnosis via radiology. The article goes into details in respect of differential diagnosis criteria, particularly within the frame of tumour diagnosis. Differentiation can be effected with the help of findings in peritoneal carcinosis and intestinal changes induced by radiation.     

Correlation of structure and function in the chicken lower intestine (coprodeum): a review

The rapid manual ParaSight-F test of Plasmodium falciparum malaria, an antigen capture test for detecting trophozoite-derived histidine rich protein-2 (PF HRP-2), is simple to perform and provides a definite diagnosis within 10 minutes. During an operational trial at health centers and mobile malaria units where microscopical diagnosis is not available and using defined symptom screening criteria, 3,361 subjects were tested yielding 618 positives (18.4%) for PF-HRP-2 by ParaSight-F. Microscopic examination of the same subjects by thick blood film examined 7 days later at a malaria clinic showed 578 falciparum, and 349 vivax and mixed infection (F+V) 41. The technology proved highly effective in detecting falciparum malaria at the peripheral levels where access to malaria laboratory services are difficult, thus allowing immediate administration of a complete course of treatment in the absence of a microscopic examination.     

Growth of sea cucumber collagen fibrils occurs at the tips and centers in a coordinated manner

Collagen fibrils are the principle source of mechanical strength in the mutable dermis of the sea cucumber Cucumaria frondosa. To obtain information about the mechanism by which collagen molecules self-assemble into fibrils, we have isolated single intact fibrils with lengths in the range 14-444 microm. These fibrils have been studied by scanning transmission electron microscopy, yielding data that show how cross-sectional mass, and hence the number of molecules in the cross-section, depend on axial location. In an individual fibril, the two ends always display similar mass distributions. The two tips of each fibril must therefore maintain identity in shape and size throughout growth. The linear relationship between cross-sectional mass and distance from the adjacent end shows that a growing tip is (like the tip of a vertebrate collagen fibril) paraboloidal in shape. Comparison of data from many different fibrils, over a wide range of lengths, however, revealed that the paraboloidal tip becomes blunter as the fibril grows in length. In contrast to vertebrate fibrils, those from C. frondosa do not have a central shaft region of constant cross-sectional mass. Rather, the cross-sectional mass increases to a maximum in the center of each fibril. The maximum cross-sectional mass of the fibrils increases exponentially with increasing fibril length. The centrosymmetry, the paraboloidal shape of the tips, and the hyperbolic increase in maximum cross-sectional mass with fibril length, is evidence for a co-ordinated regulation of length and diameter, which differs from the kind of regulation that gives rise to collagen fibrils in vertebrates (chickens and mice).     

Eicosanoid biosynthesis in an advanced deuterostomate invertebrate, the sea squirt (Ciona intestinalis)

The eicosanoid generating potential of tunic, branchial basket, intestine, ovary and tadpole larvae from the sea squirt, Ciona intestinalis, was examined using a combination of reverse phase high performance liquid chromatography, gas chromatography-mass spectrometry and enzyme immunoassay. All organs examined synthesized the lipoxygenase products 12-hydroxyeicosapentaenoic acid (12-HEPE) and 8-HEPE implying that both 8- and 12-lipoxygenase activity are widely distributed in this species. In addition, tunic and branchial basket generated significant amounts of 8,15-diHEPE and smaller amounts of 8,15-dihydroxyeicosatetraenoic acid (8,15-diHETE), while tunic alone generated small amounts of conjugated tetraene-containing material with a UV chromophore and mass ion characteristic of a lipoxin-like compound. The broad range lipoxygenase inhibitors, esculetin and nordihydroguaiaretic acid, both caused a significant dose dependent inhibition of 12-HEPE and 8,15-diHEPE biosynthesis in tunic, while the specific 5-lipoxygenase inhibitor, REV-5901, and the specific 5-lipoxygenase activating protein inhibitor, MK-866, had no observable effect on the lipoxygenase profile of this tissue. Tunic, branchial basket, intestine and ovary all generated significant amounts of prostaglandin (PG) E and PGF immunoreactive material and smaller amounts of thromboxane B immunoreactive material as measured by enzyme immunoassay. The non-specific cyclooxygenase (COX) inhibitor, indomethacin, the selective COX-1 inhibitors, resveratrol and valerylsalicylate, and the specific COX-2 inhibitors, NS-398, etolodac and DFU (5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl) phenyl-2(5H)-furanone) all caused a significant dose dependent inhibition of the biosynthesis of PGE immunoreactive material. However, the specific COX-2 inhibitors were most effective, perhaps implying that a COX-2-like enzyme may be present in this species.     

Polarity of epithelial cells of the liver. Cellular and molecular mechanisms, and pathologic changes

Smooth muscle contraction is primarily regulated not only by changes in cytosolic Ca2+ concentrations ([Ca2+]i) but also by changes in the force/[Ca2+]i ratio. The use of membrane-permeabilization technique facilitated demonstration of an increase in the level of force at constant [Ca2+]i (Ca2+ sensitization). It was clarified that Rho-associated kinase (Rho-kinase) is a novel mediator of Ca2+ sensitization of the smooth muscle contraction, by introducing the recombinant catalytic domain of Rho-kinase into the cytosol of vascular smooth muscle permeabilized with beta-escin. This review article focuses on novel mechanisms, by which activation of receptor-coupled G-protein(s) increases Ca2+ sensitivity of the contractile apparatus in smooth muscle: Rho-kinase and protein kinase C.     

Enteric coccidia (Apicomplexa) in the small intestine of the northern spotted owl (Strix occidentalis caurina)

Sporulated oocysts (mean dimensions = 13.0 x 10.8 microns) and sporocysts (11.3 x 5.5 microns) of a coccidian resembling Frenkelia sp. or Sarcocystis sp. were present in the lamina propria of the small intestine of a naturally-infected northern spotted owl (Strix occidentalis caurina) collected near Medford, Oregon (USA). Dimensions of these oocytes and sporocysts appear to be considerably smaller than those from other sarcocystid species with avian definitive hosts. Additionally, numerous developmental stages and unsporulated oocysts (mean dimensions 22.8 x 17.8 microns) of a possible species of Isospora also were observed in the intestinal epithelium. This constitutes the first report of enteric coccidia from spotted owls. Neither parasite appeared to cause the death of the bird.     

Eosinophilia and sarcoma of the small intestine (author's transl)

Eosinophilia occurs as a concomitant sign in multiple diseases of different etiology. Especially its occurrence in malignant diseases is still unknown. In three cases of primary sarcoma of the small intestine a constant eosinophilia is described. In one case this pronounced eosinophilia was present long before the final diagnose was made. The eosinophilic count was highest in the presence of clear symptoms of obstruction and normalized after surgical removal of the tumor. In one case a relapse was indicated by an increasing eosinophilic count. It is difficult to diagnose the sarcoma of the small intestine especially in its early stage because of the lack of typical symptoms. Concerning the described cases it is therefore pointed out, that in connection with undifferentiated abdominal complaints an obscure eosinophilia should be considered to be due to sarcomatous small bowel disease.     

Active transport mechanisms in the intestinal mucosa of the small intestine and their functional disorders

The efficacy of an anion-exchange gel, Secholex, as a hypocholesterolemic agent was assessed in 46 patients in 4 different studies and the effects were compared with those of cholestyramine. All patients had severe Type II-a or II-b hyperlipoproteinemia. In short-term metabolic studies Secholex (15 g/day) and cholestyramine (16 g/day) decreased serum cholesterol levels and increased total fecal sterol output and serum methyl sterol concentration to a similar extent, but cholestyramine was more effective than Secholex in increasing fecal bile acid excretion. In crossover studies, the two drugs appeared to be equally effective in lowing serum cholesterol levels but the patients mostly preferred Secholex. Twenty patients were treated with Secholex over a two-year period. The average decrease in serum cholesterol levels from the mean pretreatment value of 406 mg/100 ml was 15% during the first year, and 13% during the second year. In 5 patients the serum cholesterol was permanently lowered by more than 20% (good responders), while in 7 patients the average reduction of serum cholesterol level during Secholex administration was less than 10% (non-responders). The serum triglyceride level was slightly decreased by Secholex in Type II-b patients but was unaltered in Type II-a patients. At the end of the treatment period, serum iron and vitamin B12 levels were normal but the serum folic acid concentration was reduced in eight of 20 patients. A dose--response study indicated that a similar cholesterol-lowering effect was obtained with daily doses of 9 and 15 g of Secholex. It is concluded that Secholex is a relatively safe drug which effectively reduces serum cholesterol levels in two-thirds of patients with severe hypercholesterolemia.     

Peroxisome proliferator-activated receptor (PPAR): structure, mechanisms of activation and diverse functions

The structurally diverse xenobiotic peroxisome proliferators (PPs) increase the number of peroxisomes per cell and the levels of several enzymes, and cause hepatomegaly, often leading to hepatocarcinogenesis in a species- and tissue-specific manner. The deadlocked problems of the molecular mechanism of PP action and its physiological meanings have begun to be understood through cDNA cloning of a PP-activated receptor (PPAR). PPAR, a member of the steroid/thyroid/vitamin superfamily of nuclear receptors, has isoforms and differentially heterodimerizes with other nuclear receptors, providing potential mechanisms not only for species- and tissue-specific actions but also for diverse actions of PPs. Recent findings related to PPAR are summarized, and its possible role in lipid metabolism and involvement in PP-induced hepatocarcinogenesis are discussed.