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Description of a marine peritrichous ciliate, Pseudovorticella sinensis n. sp. (Ciliophora, Peritrichia) from China.

A new marine peritrich ciliate, Pseudovorticella sinensis n. sp. was isolated from a shrimp-farming pond in the littoral area of Qingdao, China. The morphology, infraciliature, and silverline system were studied based on living and silver-impregnated specimens. This species is characterized by (1) an elongated bell-shaped body that measures 50-60 x 35-45 microm in vivo, (2) one large, ventrally located contractile vacuole, and (3) a pellicle covered by a layer of transparent, cortical vesicles. The number of transverse silverlines from the peristomial area to the aboral ciliary wreath is 26-32, and from the aboral ciliary wreath to the scopula is 12-15. The stalk measures about 160-250 microm long x 5-6 microm wide. The spasmoneme has one row of conspicuous thecoplasmic granules, which are about 0.8 microm in diameter.

Animals↗

Dexiotrichides pangi n. sp. (protozoa, ciliophora, scuticociliatia), a new marine ciliate from the north China sea.

The morphology, infraciliature, and silverline system of a new marine scuticociliate, Dexiotrichides pangi n. sp. were investigated. The new species is characterized by: size about 45-65 x 20-25 microm in vivo with kidney-like body shape and obliquely truncated semicircle-shaped apical plate; cytostome at bottom of conspicuously depressed oral cavity, which is located at the cell equatorial level; paroral membrane extending anteriorly to membranelle 3; scutica multi-rowed; 33-38 somatic kineties; contractile vacuole near ventral side and subcaudally positioned, opening at posterior end of somatic kinety 3; one oval macronucleus and one small micronucleus; caudal cilium positioned in a small pouch; marine habitat. Based on the data obtained, an improved diagnosis for the genus Dexiotrichides is suggested: body with circular cross-section and conspicuous cilia-free apical plate; buccal cavity conspicuously depressed with cytostome located near or at equatorial level; three membranelles transversely orientated each with 2-3 rows; paroral membrane zigzaging structure, extending to about half of the length of buccal field; multi-rowed scutica; somatic kinety one strongly shortened and terminating anteriorly at posterior end of buccal field; basal bodies in equatorial region arranged usually in circular pattern, while in the anterior portion of somatic kinety 2, basal bodies characteristically in pairs and separated from the posterior part of kinety 2; one caudal cilium.

Animals↗

Holosticha hamulata n. sp. and Holosticha heterofoissneri Hu and Song, 2001, two urostylid ciliates (protozoa, ciliophora) from intertidal sediments of the yellow sea.

Two marine urostylid ciliates, Holosticha hamulata n. sp. and Holosticha heterofoissneri Hu and Song, 2001, were investigated using live observation and protargol impregnation. Both species were isolated from Korean intertidal sediments of the Yellow Sea. Holosticha hamulata measures about 150 x 25 mum in vivo, and is characterized by a tripartite body shape with a narrow head, an inflated trunk, and a tail that distally projects ventrally forming a hook-like structure. It is the characteristic body shape that distinguishes H. hamulata distinctly from congeners. Holosticha hamulata differs from H. heterofoissneri, possibly the nearest relative, also by the location of the contractile vacuole (ahead of mid-body versus near posterior body third) and the configuration of the macronucleus (on average, 33 scattered nodules assuming a Y-shape versus 17 nodules that may form a U shape). The average number of the macronuclear nodules is a pronounced feature showing great consistency in populations of each species. However, their arrangement is variable in H. heterofoissneri where the nodules are basically scattered or connected by fine fibers forming an elongate U-shape. The location of the contractile vacuole as a taxonomic feature is discussed and a dichotomous key to the species of Holosticha sensu stricto is provided.

Animals↗

Descriptions of Protospathidium serpens (Kahl, 1930) and P. fraterculum n. sp. (Ciliophora, Haptoria), two species based on different resting cyst morphology.

Protospathidium serpens (Kahl, 1930) is frequent in semiterrestrial and terrestrial habitats worldwide. Conventionally, all populations are considered as conspecific because they have very similar overall morphologies and morphometrics. We studied in detail not only the morphology of the vegetative cells but also the resting cysts using live observation, protargol impregnation, and scanning electron microscopy. These revealed a cryptic diversity and biogeographic pattern in details of the dorsal brush and cyst wall morphology. The cyst wall is spiny in the Austrian specimens, while smooth in the South African and Antarctic populations. Accordingly, P. serpens consists of at least two species: P. serpens (with spiny cyst wall) and P. fraterculum n. sp. (with smooth cyst wall); the latter is probably composed of two distinct taxa differing by the absence (South African)/presence (Antarctic) of a monokinetidal bristle tail in brush row 3, the number of dikinetids comprising brush row 1 (seven versus three), and the total number of brush dikinetids (29 versus 17). Protospathidium serpens is neotypified with the new population from Austria. The significance of resting cyst morphology is discussed with respect to alpha-taxonomy and overall ciliate diversity.

Animals↗

The apertospathulidae, a new family of haptorid ciliates (protozoa, ciliophora).

We studied the morphology of three rare haptorid ciliates, using live observation and silver impregnation: Apertospathula verruculifera n. sp., Longispatha elegans n. gen., n. sp., and Rhinothrix porculus (Penard, 1922) n. gen., n. comb. Simple ethanol fixation (50-70%, v/v) is recommended to reveal the ciliary pattern of "difficult" ciliates, such as R. porculus, by protargol impregnation. The three genera investigated have a distinct feature in common, viz., a lasso-shaped oral bulge and circumoral kinety, where the right half is slightly to distinctly longer than the left and the circumoral kinety is open ventrally. Thus, they are united in a new spathidiid family, the Apertospathulidae n. fam., which probably evolved from a Bryophyllum-like ancestor by partial reduction of the oral bulge and circumoral kinety. Apertospathula verruculifera has a wart-like process, the palpus dorsalis, at the anterior end of the dorsal brush. The right branch of the circumoral kinety is only slightly longer than the left one. Longispatha elegans has a straight oral bulge and circumoral kinety, the right branch of which extends to the posterior end of the body while the left branch ends in the anterior third of the body. Rhinothrix porculus, a curious ciliate with a snout-like dorsal elongation of the oral bulge, the palpus oralis, has a highly characteristic ciliary pattern: the oral pattern is as in Longispatha, but the bulge and circumoral kinety extend spirally to the posterior end of the body while the somatic kineties course meridionally. This is achieved by inserting some shortened kineties in the curves of the oral bulge.

Animals↗

Two new species of Epistylis (Ciliophora: Peritrichida) on the blue crab (Callinectes sapidus) in the Gulf of Mexico.

Two epibiotic peritrichs infested the blue crab, Callinectes sapidus, from the Gulf of Mexico, Mississippi, USA. Epistylis callinectes n. sp. was isolated from the epipods of maxillipeds, bases of gill-cleaning setae, and gills, and Epistylis clampi n. sp. was isolated from the exterior surfaces of the exoskeleton. Epistylis callinectes has short, symmetrically and dichotomously branched stalks; its zooid is elongate ovoid and conspicuously longer than the individual stalk branches, measuring 40-57 (49) x 18-33 (26) microm in vivo and containing a thick, undivided peristomial lip (PL). It has a single contractile vacuole and a transverse horseshoe-shaped macronucleus. Its haplokinety (H) and polykinety (Po) complete one and one-half circuits on the peristome before entering the infundibulum. There is a distal kinetal fragment present at the distal end of both the H and Po. Epistylis callinectes has 48-70 transverse silverlines from the oral area to the trochal band (TB) and 19-26 from the TB to the scopula. Epistylis clampi has long, asymmetrically, and dichotomously branched stalks. Its zooid is elongate vase-shaped, measuring 35-64 (48) x 21-30 (27) microm in vivo and with a thick, transversely folded PL. The stalks supporting zooids are unequal in length. Its zooid has a single contractile vacuole and a transverse horseshoe-shaped macronucleus occurs in the upper half of the body. Its H and Po complete approximately one circuit around the peristome before entering the infundibulum. There is a distal kinetal fragment present at the distal end of both the H and Po. This species has 71-112 transverse silverlines from the peristome to the scopula.

Animals↗

Taxonomic characterization of two marine peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp. (Ciliophora: Peritrichia), from North China.

The morphology, infraciliature, and silverline system of two new peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp., have been investigated based on both living and silver-impregnated specimens. Partial sequence of 18S-ITS1-5.8S rDNA of Z. pararbuscula is also determined in order to compare it with the closely related congener, Zoothamnium arbuscula. Zoothamnium pararbuscula can be distinguished from its close form Z. arbuscula by the different habitats, the appearance of the main stalk, the position of the contractile vacuole, and the information derived from 18S-ITS1-5.8S rDNA sequence analysis. Pseudovorticella clampi n. sp. is distinguished from its congeners by its body shape and size, pellicle granules, habitat, and number of transverse silverlines.

Animals↗

Morphology and infraciliature of three species of Metaurostylopsis (Ciliophora, Stichotrichia): M. songi n. sp., M. salina n. sp., and M. marina (Kahl 1932) from sediments, saline ponds, and coastal waters.

Two new urostylid ciliates, Metaurostylopsis songi n. sp. and Metaurostylopsis salina n. sp. and Metaurostylopsis marina (Kahl 1932) are investigated using live observation and protargol impregnation. These species were isolated in Korea from intertidal sediments, saline ponds, and coastal waters. Metaurostylopsis songi is in vivo about 120 microm x 25 microm, has a slenderly ellipsoidal body, colorless cortical granules in rows on ventral and dorsal body sides, about 54 macronuclear nodules, 28-47 adoral membranelles, five frontal, two or three frontoterminal and six or seven transverse cirri, and 9-12 midventral cirral pairs followed posteriorly by 1-3 single cirri. In vivo M. salina is about 60 microm x 25 microm, has a pyriform body, colorless cortical granules irregularly arranged, about 45 macronuclear nodules, 18-23 adoral membranelles, three frontal, three to five frontoterminal and two to five transverse cirri, and four or five midventral cirral pairs followed posteriorly by five to seven single cirri. Both species have three marginal cirral rows on each body side and 3 long dorsal kineties. The Korean specimens of M. marina match the Chinese population in all main features. Metaurostylopsis songi differs from M. marina by the more slender body, the number of frontal cirri (invariably five vs. four), and the arrangement of cortical granules (in rows on dorsal and ventral cortex vs. only along dorsal kineties and anterior body margin). Metaurostylopsis salina differs from its congeners by the distinctly smaller size, the pyriform body shape, the scattered cortical granules (vs. in rows), and number of frontal cirri. It differs from M. marina also by the number of midventral cirral pairs (four or five vs. seven to 11).

Animals↗

Growth-phase dependent substrate adhesion in Crithidia fasciculata.

Crithidia fasciculata is a trypanosomatid flagellate that parasitizes several species of mosquito. Within the alimentary tract of its host, C. fasciculata exists in two forms: one is a non-motile form, attached in clusters to the lining of the gut, the other a more elongated form swimming freely in the gut lumen. We have developed an in vitro culture system that reproduces the appearance of these two distinct morphological forms. Using two different cultivation methods, shaking and stationary incubations, we have demonstrated that adherence phenotypes are growth-phase dependent. Organisms in the logarithmic phase of growth possess the ability to adhere to substrates; this ability is lost when the organism enters a stationary growth phase. Parasite adherence was independent of cultivation method or substrate. Furthermore, adherent forms of Crithidia maintained their adhesive properties following their removal from substrates. Our data reveal a growth-phase-regulated process of cell attachment that may influence the transmission and dissemination of this parasitic flagellate.

Animals↗

Splenic function in chronic myelogenous leukemia.

Spleen function was evaluated by measurement of the clearance of autologous heat-damaged 99mTc-labelled erythrocytes from the circulation and into the spleen and the enumeration of pitted erythrocytes by interference contrast microscopy, and the spleen area was determined by scintillation scanning. All measurements were performed on 12 patients with chronic myelogenous leukemia and compared with 10 controls with apparently normal spleens, 6 splenectomized subjects and 9 patients with a reactive splenomegaly. Patients with CML had spleen function test results similar to normal controls in spite of having enlarged spleens whose projection area did not differ from that of the patients with reactive splenomegaly. Thus, patients with CML have a decreased spleen function per unit volume and signs of splenic hypofunction in the peripheral blood. The reduction of spleen function per unit volume in CML is the result of a severe decrease of the splenic blood perfusion which could result from the combined effects of the myeloid metaplasia and the increased whole-blood viscosity due to high white-cell counts.

Adolescent↗

Effect of cytochalasin B and dihydrocytochalasin B on invasiveness of entero-invasive bacteria in HEp-2 cell cultures.

The effect of cytochalasin B (CB) and di-hydrocytochalasin B (H2CB) on the invasiveness of Salmonella typhimurium, Shigella flexneri and Yersinia enterocolotica serotype 0:3 in HEp-2 cell cultures was examined. The intra-cellular and extra-cellular bacteria were identified by a combination of Nomarski interference contrast microscopy and UV incident light microscopy applied on the same light microscope. Pre-treatment of cells with CB and H2CB inhibited the uptake of S. typhimurium and S. flexneri in the HEp-2 cell cultures. The effect was time and dose dependent. On the other hand, the drugs did not influence the invasiveness of Y. enterocolitica. The results indicate that activity of cellular actin micro-filaments is essential for invasiveness of S. typhimurium and S. flexneri.

Cells, Cultured↗

Effects of gonadotrophins on the cumulus oophorus of isolated rat Graafian follicles.

Graafian follicles were extirpated before the endogenous LH surge on the day prior to ovulation from PMS-injected immature rats. They were incubated in chemically defined medium in presence or absence of gonadotrophins for 4-10 h. After incubation the oocyte-cumulus complexes were recovered from the follicles and one group inspected by Nomarski interference contrast microscopy and one group was placed in saline containing hyaluronidase. In hormone-free medium both the oocyte and the cumulus cells remained morphologically unchanged: a dictyate oocyte surrounded by a compact mass of cumulus cells. Hyaluronidase did not detach cells from the cumulus structure. However, in presence of LH or FSH morphological changes developed in both cell types: the oocyte resumed meiosis as revealed by germinal vesicle breakdown and polar body formation; the cumulus structure became dispersed and embedded in a viscous matrix. The individual cumulus cells displayed pseudopodia-like processes of the cell surface. Treatment with hyaluronidase resulted in detachment of cells from the cumulus. The effects of LH described on the oocyte and cumulus cells were unimpaired in presence of cyanoketone or aminoglutethimide although these agents blocked the follicular relase of progesterone, estradiol and androstenedione as revealed by RIA. The results indicate that a close functional relationship exists between the two cell types of the cumulus oophorus, the oocyte and the cumulus granulosa cells. Both cell types are affected directly or indirectly by gonadotrophins and characteristical morphological changes develop synchronously in both cell types.

Aminoglutethimide↗

Decreasing hair cell counts in aging humans.

Deterioration of balance with advancing age is a well-known fact of life. Some investigators have reported a 50% prevalence of dizziness in the elderly. Clinically, progressive dysequilibrium of aging presents as gradually worsening balance due to age-related decline in function of the peripheral vestibular system, central nervous system, vision, and musculoskeletal system. Vestibular function testing has shown clear evidence of age-related changes in peripheral and central sites. Histopathologic changes in the vestibular sensory organs include progressive hair cell degeneration, otoconial degeneration in the otolith organs, and decreasing number of Scarpa's ganglion neurons. Recently, a new quantitative method of assessing vestibular otopathology has been described, utilizing Nomarski differential interference contrast microscopy. This technique has been applied to 67 human temporal bones of individuals from birth to age 100 to create a normative database of total, type I, and type II hair cell counts as a function of age. Results show a highly significant continuous decrease in all counts from birth to age 100, best fit by a linear regression model. Type I hair cell counts in all three semicircular canal cristae decrease at a similar rate, significantly faster than the degeneration observed in type I hair cells of the maculae. Type II hair cell counts decline at the same rate for all 5 sensory epithelia. These normative data provide the basis for comparisons to hair cell counts made in temporal bones from subjects with known vestibular disorders. They also provide a basis for drawing correlations between vestibular function testing and vestibular otopathology.

Aged↗

Electrical membrane responses to secretagogues in parietal cells of the rat gastric mucosa in culture.

Fragments of the gastric fundus of 6-8-day-old rats were maintained in tissue culture. From the explant, adhered to a plastic substrate, epithelial cells migrated and developed to form a monolayer colony. Histological and histochemical studies as well as indirect immunofluorescence studies using anti-parietal cell antibodies testified to the presence of parietal cells in the monolayer during the first week. These parietal cells were distinguished by their vesicular cytoplasmic structures using phase-contrast or differential interference-contrast microscopy. Acridine Orange, an optical probe of H+ accumulation, was taken up preferentially by these parietal cells, exhibiting orange fluorescence within the cells on the third day of culture, in response to stimulation with gastrin, histamine and carbachol. The resting potential of these cultured parietal cells was about -20 mV. On day 2-4 of culture, the cell membrane became hyperpolarized (up to -30 to -40 mV) in response to gastrin, carbachol or histamine in the presence of isobutylmethyl-xanthine (IMX). During hyperpolarization, the membrane resistances decreased significantly. The amplitude and the polarity of secretagogue-induced responses were found to be dependent on the extracellular concentration of K+ (but not Na+ and Cl-). The carbachol-induced responses were inhibited by atropine but not curare. The responses induced by histamine plus IMX were blocked by cimetidine but not pyrilamine. Neither atropine nor cimetidine affected the gastrin-evoked responses. It is concluded that rat parietal cells have separate receptors for acetylcholine (muscarinic), gastrin and histamine (H2), and that an increase in the membrane permeability to K+ is closely associated with the responses of these receptors under these in vitro conditions.

Animals↗

Contact deformation of liposome in the presence of osmosis.

The role of osmotic pressure on the geometry of adherent liposome remains an intricate question in the mechanics of supramolecular structures. In this study, confocal reflection interference contrast microscopy in combination with cross-polarized microscopy was applied to probe the geometry of deformed liposome on fused silica substrates through the determination of a vesicle-substrate separation profile. In parallel, a theoretical model which describes the large deformation of the lipid bilayer membrane under both out-plane bending and in-plane shear forces is developed. Then, the global deformation geometry of the adherent liposome is rigorously compared with our experimental data. It is shown that the adhesion contact area increases in dimension, the liposome volume decreases, and the vesicle height decreases under the reduced osmotic pressure. The coupling of experimental data and a modified theoretical framework of the adherent liposome provides a more explicit result in comparison with previous studies and demonstrates the possibility of modeling the change of liposome mechanics under the influence of osmosis.

Adsorption↗

Tensile mechanical properties of three-dimensional type I collagen extracellular matrices with varied microstructure.

The importance and priority of specific micro-structural and mechanical design parameters must be established to effectively engineer scaffolds (biomaterials) that mimic the extracellular matrix (ECM) environment of cells and have clinical applications as tissue substitutes. In this study, three-dimensional (3-D) matrices were prepared from type I collagen, the predominant compositional and structural component of connective tissue ECMs, and structural-mechanical relationships were studied. Polymerization conditions, including collagen concentration (0.3-3 mg/mL) and pH (6-9), were varied to obtain matrices of collagen fibrils with different microstructures. Confocal reflection microscopy was used to assess specific micro-structural features (e.g., diameter and length) and organization of component fibrils in 3-D. Microstructural analyses revealed that changes in collagen concentration affected fibril density while maintaining a relatively constant fibril diameter. On the other hand, both fibril length and diameter were affected by the pH of the polymerization reaction. Mechanically, all matrices exhibited a similar stress-strain curve with identifiable "toe," "linear," and "failure" regions. However the linear modulus and failure stress increased with collagen concentration and were correlated with an increase in fibril density. Additionally, both the linear modulus and failure stress showed an increase with pH, which was related to an increasedfibril length and a decreasedfibril diameter. The tensile mechanical properties of the collagen matrices also showed strain rate dependence. Such fundamental information regarding the 3-D microstructural-mechanical properties of the ECM and its component molecules are important to our overall understanding of cell-ECM interactions (e.g., mechanotransduction) and the development of novel strategies for tissue repair and replacement.

Animals↗