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PubMed1995

The fate of the Golgi apparatus and the endoplasmic reticulum during lens fiber cell differentiation.

PURPOSE: To establish the fate of the Golgi apparatus and the endoplasmic reticulum (ER) during lens fiber differentiation. METHODS: Organelles were visualized by confocal or electron microscopy. For fluorescence microscopy, organelles were labeled with fluorescent probes or antibodies raised against organelle-resident proteins. The cytoplasmic volume was reconstructed from optical sections using volume rendering techniques. RESULTS: The Golgi apparatus was located apically in epithelial cells. In the annular pad, Golgi elements were transformed into ribbon-like structures running parallel to the long axes of the cells. Toward the lens equator, the Golgi apparatus fragmented. In the lens fibers, the Golgi apparatus was detected only in the superficial cells. The ER was present as vesicular or tubular elements in both epithelial and cortical fiber cells, and ER probes co-labeled the nuclear membrane and revealed that the ER and nuclei disappeared coincidentally in the deep cortex. Using a lipophilic dye and volume rendering, the relationships between organelles could be evaluated in three dimensions. CONCLUSIONS: The Golgi apparatus was not a prominent organelle in differentiating lens fibers. In contrast, the ER was more abundant and extended to the edge of the organelle-free region, where it was degraded along with the nuclei and mitochondria.

Amino Acid Sequence↗
PubMed2000

A novel apparatus for rat in vivo evaluation of dry powder formulations for pulmonary administration.

The lungs have attracted increasing attention as a site for administration of drugs, including macromolecules that are poorly absorbed from the intestine. There have been a number of basic studies in which peptide solutions were administered to experimental animals via the lungs. Although there have been several studies of pulmonary peptide absorption from dry powder formulations, a simpler and more inexpensive apparatus for administration of dry powders would enhance rapid screening of the formulations. In this study, we developed a simple apparatus to disperse dry powders. The apparatus has two 3-way stopcocks; one allows dispersal of powders at a constant pressure and airflow, and the other allows rats to breathe before and after administration. Dry powders of fluorescein (FL) and FITC-dextran (FD4) were manufactured by the spray-drying technique. The effects of operating conditions on the absorption of these model drugs were examined in rats. The C(max) for FL from dry powder was lower than that from solution and mean residence time was extended, suggesting that dissolution was the rate-determining step for FL absorption from dry powder. For FD4, the rate of absorption may not be regulated by dissolution but by epithelial transport. Absorption of insulin from spray-dried powder via the rat trachea was investigated using this apparatus. Intratracheally administered spray-dried insulin powder decreased plasma glucose level to a greater extent than spray-dried insulin solution administered via the same route. Thus, the apparatus is simple, inexpensive, and useful for rapid screening of dry powder formulations.

Absorption↗
PubMed2002

Regulation of flagellar dynein by the axonemal central apparatus.

Numerous studies indicate that the central apparatus, radial spokes, and dynein regulatory complex form a signaling pathway that regulates dynein activity in eukaryotic flagella. This regulation involves the action of several kinases and phosphatases anchored to the axoneme. To further investigate the role of the central apparatus in this signaling pathway, we have taken advantage of a microtubule-sliding assay to assess dynein activity in central apparatus defective mutants of Chlamydomonas. Axonemes isolated from both pf18 and pf15 (lacking the entire central apparatus) and from pf16 (lacking the C1 central microtubule) have reduced microtubule-sliding velocity compared with wild-type axonemes. Based on functional analyses of axonemes isolated from radial spokeless mutants, we hypothesized that inhibitors of casein kinase 1 (CK1) and cAMP dependent protein kinase (PKA) would rescue dynein activity and increase microtubule-sliding velocity in central pairless mutants. Treatment of axonemes isolated from both pf18 and pf16 with DRB, a CK1 inhibitor, but not with PKI, a PKA inhibitor, restored dynein activity to wild-type levels. The DRB-induced increase in dynein-driven microtubule sliding was inhibited if axonemes were first incubated with the phosphatase inhibitor, microcystin. Inhibiting CK1 in pf15 axonemes, which lack the central pair as well as PP2A [Yang et al., 2000: J. Cell Sci. 113:91-102], did not increase microtubule-sliding velocity. These data are consistent with a model in which the central apparatus, and specifically the C1 microtubule, regulate dynein through interactions with the radial spokes that ultimately alter the activity of CK1 and PP2A. These data are also consistent with localization of axonemal CK1 and PP2A near the dynein arms.

Alleles↗
PubMed2001

Cardiac transplantation following a 24-h preservation using a perfusion apparatus.

BACKGROUND: We developed a new apparatus for heart preservation and have already reported successful transplantation following 12 h of preservation using this apparatus. The efficacy of coronary perfusion with an oxygenated Celsior solution was investigated through transplantation following 24 h of preservation using the apparatus. MATERIALS AND METHODS: After being harvested, grafts were preserved with a combination of immersion in a 4 degrees C Celsior solution and perfusion with an oxygenated Celsior solution using the apparatus in the coronary perfusion (CP) group and simply immersed in a 4 degrees C Celsior solution in the simple immersion(SI) group. beta-Adenosine triphosphate (beta-ATP), phosphocreatine (Pcr), and inorganic phosphate (P(i)) levels and myocardial pH (pH(i)) were measured immediately after the heart was excised and at 12 and 24 h after preservation. Following preservation, orthotopic transplantation was performed. Cardiac function was measured 2 h after weaning from cardiopulmonary bypass (CPB). RESULTS: beta-ATP/P(i), Pcr/P(i), and pH(i) levels were significantly higher in the CP group than in the SI group at 12 and 24 h after preservation. Four of six animals in the CP group and two of six in the SI group were successfully weaned from CPB. The recovery rates of cardiac function were better in the CP group than in the SI group. CONCLUSION: Twenty-four hours of heart preservation may be possible with a combination of immersion in a 4 degrees C Celsior solution and perfusion with an oxygenated Celsior solution using the perfusion apparatus.

Animals↗
PubMed1985

Relationships between synaptic junctions, puncta adhaerentia and the spine apparatus at neocortical axo-spinous synapses. A serial section study.

A total of 80 cotical axo-spinous synaptic junctions were reconstructed from serial sections and about 100,000 were analyzed in single sections. Special attention was paid to the occurrence of puncta adhaerentia associated with perforated, annulate or horseshoe-shaped (= complex) synaptic junctions and to the presence and proximity of the spine apparatus. Further evidence is presented that the spine apparatus has no relationship to simple (round or oval) synaptic specializations, but is present in association with at least 91% of complex junctions. The spine apparatus points towards the punctum adhaerens which in at least 71% of cases seems to be an integral part of the complex synapse. Direct continuity was found between the dense material of the spine apparatus and the punctum adhaerens. It is suggested, in accordance with other recent studies, that expansion of the synaptic active zone occurs by the addition and transformation of puncta adhaerentia. The spine apparatus may participate in this dynamic process as a possible donor of specific postsynaptic proteins.

Adult↗
PubMed2006

Structural and immunocytochemical characterization of the Ginkgo biloba L. sperm motility apparatus.

Ginkgo biloba and the cycads are the only extant seed plants with motile sperm cells. However, there has been no immunocytochemical characterization of these gametes to determine if they share characteristics with the flagellated sperm found in bryophytes and pteridophytes or might give clues as to the relationships to nonflagellated sperm in all other seed plants. To determine characteristics of proteins associated with the motility apparatus in these motile sperm, we probed thin sections of developing spermatogenous cells of Ginkgo biloba with antibodies to acetylated and tyrosinated tubulin and monoclonal antibodies that recognize mammalian centrosomes and centrin. The blepharoplast that occurs as a precursor to the motility apparatus consists of an amorphous core, pitted with cavities containing microtubules and a surface studded with probasal bodies. The probasal bodies and microtubules within the blepharoplast cavities are labeled with antibodies specific to acetylated tubulin. Positive but weak reactions of the blepharoplast core occur with the centrosomereactive antibodies MPM-2 and C-9. Reactions to centrin antibodies are negative at this developmental stage. From this pre-motility apparatus structure, an assemblage of about 1,000 flagella and associated structures arises as the precursor to the motility apparatus for the sperm. The flagellar apparatus consists of a three-layered multilayered structure that subtends a layer of spline microtubules, a zone of amorphous material similar to that in the blepharoplast, and the flagellar band. Centrin antibodies react strongly with the multilayered structure, the transition zone of the flagella, and fibrillar material near the flagellar base at the surface of the amorphous material. Both the spline microtubules and all of the tubules in the flagella react strongly with the antibodies to acetylated tubulin. These localizations are consistent with the localizations of these components in pteridophyte and bryophyte spermatogenous cells, although the blepharoplast material surrounding and connecting flagellar bases does not occur in the seedless (nonseed) land plants. These data indicate that despite the large size of ginkgo gametes and the taxonomic separation between pteridophytes and Ginkgo biloba, similar proteins in gametes of both groups perform similar functions and are therefore homologous among these plants. Moreover, the presence of acetylated tubulin in bands of microtubules may be a characteristic shared with more derived non-flagellated sperm of other conifers and angiosperms.

Cell Movement↗
PubMed1990

A vertical submarine electrophoresis apparatus for polyacrylamide minigels.

A vertical submarine electrophoresis apparatus for use with minislab polyacrylamide gels is described. The design allows polyacrylamide gels to be run with the same ease and convenience that agarose gels are run with horizontal submarine apparatuses. The vertical submarine features a single buffer chamber with a restriction between the upper and the lower portions of the chamber. Acrylamide gels, cast between 9 X 10-cm glass slides, are inserted into the restriction and are completely immersed in buffer. Thus, current flows primarily through the gel itself, but some current flows through the buffer in the restriction surrounding the gel. Because water-tight separation of buffer chambers is not necessary, time-consuming and/or expensive procedures such as sealing with agarose or using fragile notched glass plates are eliminated. The apparatus can be set up to run a gel in less than 30 s. It is versatile in that gels of varying thickness (0.5, 0.8, 1.5, and 3 mm) can be run on a single apparatus. The apparatus has been used for sodium dodecyl sulfate gels, low ionic strength native gels for nucleoprotein complexes, and composite acrylamide-agarose gels.

Chemistry Techniques, Analytical↗
PubMed1984

The ciliary basal apparatus is adapted to the structure and mechanics of the epithelium.

The basal apparatuses which anchor the gill cilia in Branchiostoma lanceolatum (Pallas) and the actinopharynx cilia in Calliactis parasitica (Couch) are similar in structure. In C. parasitica the pharynx epithelium and the basal apparatuses are flexible. The basal apparatuses, however, bend in only one direction. This mechanism may permit epithelial flexibility whilst maintaining a similar basal orientation between cilia. In B. lanceolatum the ciliated gill epithelia are mechanically stable but the epithelial surfaces are curved. The basal apparatuses may correct for this curvature, with short rootlets between the distal centrioles (basal bodies) and the cell membranes, so that their cilia also share a common orientation. A common basal orientation between cilia is important for their coordination. The degree of coordination depends upon the function of the cilia; water-propelling cilia are more precisely coordinated than mucus-propelling cilia. Much of the structural diversity of ciliary basal apparatuses in Metazoa may be due to variation in the demands of anchoring functionally different cilia to epithelia which have different structural and mechanical properties.

Animals↗
PubMed2004

A new acceleration apparatus for the study of whiplash with human cadaveric cervical spine specimens.

The biomechanics of whiplash is often studied using cadaveric cervical spine specimens. One of the most important points in this kind of study is to create realistic loading conditions. The aim of the present project therefore was to develop an acceleration apparatus, which allows the study of whiplash with human cadaveric cervical spine specimens under as realistic loading conditions as possible. The new acceleration apparatus mainly consisted of a sled, a pneumatic acceleration unit and a railtrack and offered several unique features to create more realistic loading conditions. Among these features, the possibility to simulate the passive movements of the trunk is of capital importance. In this new apparatus, first, the general feasibility of whiplash experiments was studied, second, the reproducibility of the impacts was quantified and third, the effect of simulated movements of the trunk on accelerations and loads was examined. In the new acceleration apparatus various types of collisions could reproducibly be simulated. Simulated passive movements of the trunk strongly influenced the loading pattern of the neck. Without pivoting a steep increase of all loading parameters could be observed. This increase was less pronounced if pivoting was allowed. In conclusion, biomechanical aspects of whiplash could reproducibly be examined in the new acceleration apparatus. Due to its significant effects on the loading of the neck, pivoting of the trunk should always be taken into account in future experiments on the biomechanics of whiplash in which isolated cervical spine specimens are used.

Acceleration↗
PubMed2005

A case of abnormal findings in the course of the vertebral artery associated with an ossified hyoid apparatus. A contraindication for manipulation of the cervical spine?

OBJECTIVE: To describe a case of a simultaneous occurrence of an ossified stylohyoid ligament in a 56-year-old male cadaver and anomalies of the vertebral artery, and to consider the clinical implications for manipulative therapists. INTERVENTION AND OUTCOME: Dissection showed a simultaneous occurrence of complete developmental ossification of the left hyoid apparatus, variants of the vertebral artery, and a left superior vena cava in a 56-year-old male cadaver. DISCUSSION: Developmental variants, posttraumatic and degenerative changes of the hyoid apparatus may result in variable degrees of ossification or calcification. CONCLUSION: This unusual disorder should be considered in the differential diagnosis of facial and neck pain especially within the scope of manipulation of the upper cervical spine. Cervical spine manipulation may exacerbate existing pathological conditions of the stylohyoid apparatus, thereby irritating neurovascular structures, and induce a fracture. Developmental ossification of this apparatus might be associated with anomalies in the atlantic section of the vertebral artery which make the patient more susceptible to vertebrobasilar insufficiency. We conclude that extreme care should be taken in the presence of such an ossification to avoid trauma to the stylohyoid apparatus and maybe even because of increased vertebrobasilar risk.

Cadaver↗
PubMed2006

A computerized apparatus designed to automatically dispense, measure, and record alcohol consumption by individual members of a rhesus macaque social group: trait-like drinking across social- and single-cage conditions.

BACKGROUND: The present study tested the validity of an automated ethanol dispensing apparatus that is capable of identifying individual monkeys and precisely measuring their levels of ethanol consumption while living in a social group, and assessed individual subjects' level of consumption when alone and in social groups. METHODS: In Experiment 1, 21 rhesus macaques (Macaca mulatta) were given access for 1-h each day to the dispensing apparatus, which contained an aspartame-sweetened 8.4% (v/v) ethanol solution. Measurements of blood ethanol concentrations were taken for each subject and compared with the level of consumption recorded by the apparatus for those subjects. To examine the possibility that competition among the animals limited their access to the dispensing unit, in Experiment 2, 10 of the subjects used in Experiment 1 were singly housed to allow them to drink without interference from other monkeys. A correlation was then performed to assess the interindividual relationship between the amount of ethanol consumed in these two housing conditions. RESULTS: In Experiment 1, the volume of solution measured and recorded by the apparatus correlated positively with the true volume dispensed. Furthermore, the volume of solution reported by the computer to have been consumed by an individual subject correlated positively with blood ethanol concentrations. In Experiment 2, the volume of ethanol consumed by individual subjects in single cages correlated positively with their consumption in the social group. CONCLUSIONS: The apparatus accurately identified and measured individual patterns of ethanol consumption among socially housed animals. Additionally, individual differences in ethanol consumption remained stable across settings, as shown by the strong positive correlation between drinking in a social setting versus drinking alone. This finding may thus reflect an individual's constitutional proclivity to consume alcohol.

Alcohol Drinking↗
PubMed2002

An apparatus for color Doppler imaging in seated subjects.

PURPOSE: To easily perform reliable color Doppler imaging of the orbital vessels in seated subjects. DESIGN: Experimental study. METHODS: Experimental study. An apparatus mounting the color Doppler imaging probe through a spring plate on a slit-lamp system was devised. In 16 eyes of eight seated subjects, color Doppler imaging assessments of the central retinal artery using the apparatus and by the examiner's handheld manipulation were repeated with a 10-minute interval. The reproducibility coefficients were calculated regarding the color Doppler imaging indices, including peak systolic velocity, end diastolic velocity, and resistive index. RESULTS: The color Doppler imaging indices obtained with the apparatus showed no significant differences from those obtained by handheld manipulation. The time required for each measurement using the apparatus was shorter than that by handheld manipulation, and reproducibility coefficients were significantly smaller by about 50%. CONCLUSION: Color Doppler imaging can be easily performed with good reproducibility in seated subjects using this apparatus.

Adult↗
PubMed2002

Reoperative mitral valve replacement: importance of preservation of the subvalvular apparatus.

BACKGROUND: Preservation of the subvalvular apparatus has been demonstrated to be beneficial during first-time mitral valve replacement (MVR), but has not been fully examined in reoperative (redo) MVR. The purpose of this study was to analyze outcomes in a large cohort of redo MVR patients, focusing on the effect of subvalvular preservation on mortality. METHODS: We undertook a review of prospectively gathered data on patients undergoing MVR, with or without concomitant cardiac procedures, at our institution from 1990 to 1999. Predictors of mortality were determined by stepwise logistic regression. RESULTS: A total of 1,521 consecutive MVR patients were analyzed, of which, 513 (34%) had undergone one or more previous MV procedures. In-hospital mortality occurred in 6.9% of first-time MVR patients versus 9.0% in redo patients (p = 0.13). The number of prior MV operations ranged from one to five in redo MVR patients, with 115 patients (22% of redos) having two or more. In redo MVR patients, preservation of the native posterior subvalvular apparatus was performed in 103 patients (21%), whereas native anterior and posterior preservation was performed in 31 patients (6%). Gore-Tex neochordal construction was performed in 135 redo MVR patients (26%). Perioperative mortality occurred in 3.6% of redo MVR patients with a preserved subvalvular apparatus (native tissue and/or Gore-Tex reconstruction) versus 13.3% of redo patients without preservation (p < 0.001). Independent predictors of mortality in redo MVR patients were (in decreasing order of magnitude) failure to preserve the subvalvular apparatus, preoperative renal failure, previous stroke/transient ischemic attack, left ventricular dysfunction (left ventricular ejection fraction <40%), and urgent timing. CONCLUSIONS: Redo MVR can be performed with an acceptable risk of mortality. Although preservation of the subvalvular apparatus may increase operative complexity, we recommend subvalvular preservation in order to decrease the risk of early mortality.

Adult↗
PubMed1996

Structural and functional reconstitution of thin filaments in the contractile apparatus of cardiac muscle.

The muscle contractile apparatus has a highly ordered liquid crystalline structure. The molecular mechanism underlying the formation of this apparatus remains, however, to be elucidated. Selective removal and reconstitution of the components are useful means of examining this mechanism. In addition, this approach is a powerful technique for examining the structure and function of a specific component of the contractile system. In this study we have achieved the structural and functional reconstitution of thin filaments in the cardiac contractile apparatus. First, all thin filaments other than short fragments at the Z line were removed by treatment with gelsolin. Under these conditions no active tension could be generated. By incorporating exogenous actin into these thin filament-free fibers, actin filaments were reconstituted, and active tension, which was insensitive to Ca2+, was restored. The active tension after the reconstitution of thin filaments reached 135 +/- 64% of the original level. The augmentation of tension was attributable to the elongation of reconstituted filaments. As another possibility for augmented tension generation, we suggest the presence of an inhibitory system that was not reconstituted. In any case, the thin filaments of the cardiac contractile apparatus are considered to be assembled so as not to develop the highest degree of tension. Incorporation of the tropomyosin-troponin complex fully restored Ca2+ sensitivity without affecting maximum tension. The present results indicate that a muscle contractile apparatus with a higher order structure and function can be constructed by the self-assembly of constituent proteins.

Actins↗
PubMed2002

Effects of prior apparatus experience and novelty of testing environment on locomotor activity following MK-801.

Robust increases in locomotor activity are observed following administration of dizocilpine maleate (MK-801). The present study investigated the effects of prior apparatus experience and manipulation of the testing environment on locomotor activity following peripheral MK-801. Gerbils were given zero or nine sessions of apparatus exposure prior to testing with MK-801 (0.1 mg/kg ip) or saline. Sessions were 10 min in duration and separated by 24 h. As previously reported, naive animals treated with MK-801 were significantly more active relative to controls. Exposure to the apparatus for nine sessions resulted in a significant reduction in MK-801-induced activity, but did not alter the activity levels of control animals. To evaluate the effect of changes to the testing environment, animals previously evaluated in the familiar condition were retested in the identical apparatus relocated to a novel experimental room. MK-801-treated animals exhibited a significant increase in activity when tested in this novel environment while the locomotor activity of control gerbils was not significantly altered. The results illustrate the importance of repetitive testing and environmental changes as moderating variables in studies that evaluate locomotor activity. These data also indicate that the effects of MK-801 on activity are sensitive to prior experience with the apparatus and the novelty of the testing environment.

Animals↗
PubMed1976

Apparatus for patient-controlled administration of intravenous narcotics during labour.

The intermittent intravenous administration of narcotic analgesics had advantages over intramuscular administration, but is often impracticable. The design of a prototype apparatus is described, from which analgesic, pethidine, can be self-administered intravenously during labour. The apparatus consists of a syringe pump and control system from which the patient can demand an increment of analgesic. The minimum interval between increments and the dose of the increment are preset. A demand for analgesia is only successful when the patient passes a reaction-time test, subsequently modified. The apparatus has been used by forty-two mothers in labour. The mean total dose self-administered (2.2 mg/kg) was well within the prescribed limits, suggesting that mothers regulated their demand. The apparatus appears acceptable and safe in labour. The apparatus will probably be of value in comparative studies of analgesics.

Anesthesia, Intravenous↗
PubMed2005

A novel apparatus for non-contact measurement of heart rate variability: a system to prevent secondary exposure of medical personnel to toxic materials under biochemical hazard conditions, in monitoring sepsis or in predicting multiple organ dysfunction syndrome.

BACKGROUND: The impaired balance of the low-frequency/high-frequency ratio obtained from spectral components of RR intervals can be a diagnostic test for sepsis. In addition, it is known that a reduction of heart rate variability (HRV) is useful in identifying septic patients at risk of the development of multiple organ dysfunction syndrome (MODS). We have reported a non-contact method using a microwave radar to monitor the heart and respiratory rates of a healthy person placed inside an isolator or of experimental animals exposed to toxic materials. APPARATUS DESIGN AND TESTING: With the purpose of preventing secondary exposure of medical personnel to toxic materials under biochemical hazard conditions, we designed a novel apparatus for non-contact measurement of HRV using a 1215 MHz microwave radar, a high-pass filter, and a personal computer. The microwave radar monitors only the small reflected waves from the subject's chest wall, which are modulated by the cardiac and respiratory motion. The high-pass filter enhances the cardiac signal and attenuates the respiratory signal. In a human trial, RR intervals derived from the non-contact apparatus significantly correlated with those derived from ECG (r=0.98, P<0.0001). The non-contact apparatus showed a similar power spectrum of RR intervals to that of ECG. CONCLUSIONS: Our non-contact HRV measurement apparatus appears promising for future pre-hospital monitoring of septic patients or for predicting MODS patients, inside isolators or in the field for mass casualties under biochemical hazard circumstances.

Adult↗
PubMed2005

Fine structure of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 (Monogenea: Ancyrocephalidae).

A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus.

Animal Structures↗