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The effect of femoral lengthening on knee articular cartilage: the role of apparatus extension across the joint.

Loss of joint motion is a common complication of limb lengthening despite newer methods of incremental bone elongation. A pilot canine study has demonstrated that 30% femoral lengthening causes reproducible knee cartilage injury manifest by frank loss of cartilage substance or fibrillation. This study was undertaken to examine the potential of knee joint protection by apparatus extension to the tibia. Four dogs underwent application of a modified Ilizarov apparatus to the femur and tibia with coaxial hinges at the knee. After osteotomy, 30% lengthening was undertaken at 0.75 mm daily in three increments. At the completion of lengthening, experimental and contralateral knee joints were harvested, assessed grossly, decalcified, sagittally sectioned, and stained with safranin-O. All control joints were normal histologically. All experimental joints demonstrated a decrease in proteoglycan staining without evidence of fibrillation or necrosis. These findings suggest a protective effect of the tibial apparatus by avoiding joint compression.

Animals↗

Maximal stress test performance while wearing a self-contained breathing apparatus.

Fifteen adult male volunteers (mean age = 31.0 years; eight non-smokers and seven smokers) carried out a Bruce protocol maximal treadmill stress test under four separate conditions. These four conditions were (1) without Scott Air Pak (SAP) respirator; (2) with SAP respirator apparatus but not wearing face mask; (3) with SAP apparatus, wearing face mask in "demand" breathing mode; and (4) with SAP respirator apparatus, wearing face mask in "pressure-demand" breathing mode. The data indicate that the overriding factor in the approximately 20% decrement in work performance during conditions 2, 3, and 4 was related to the weight of the SAP respirator (15.8 kg), P less than 0.05. Only during condition 3 did eight of 15 subjects report lack of air as one reason for termination. Conditions 3 and 4 resulted in a significant decrease in TFore (2.5 degrees C) and was primarily related to the decreased Tinsp which reached a maximum decrement of 9.7 degrees C. Under all conditions the smokers' work time was significantly less than nonsmokers, P less than 0.05.

Adult↗

Human cytomegalovirus UL37 immediate-early regulatory proteins traffic through the secretory apparatus and to mitochondria.

The human cytomegalovirus (HCMV) UL36-38 immediate-early (IE) locus encodes the UL37 exon 1 (pUL37x1) and UL37 (gpUL37) regulatory proteins, which have anti-apoptotic activities. pUL37x1 shares its entire sequence, including a hydrophobic leader and an acidic domain, with the exception of one residue, with the amino terminus of gpUL37. gpUL37 has, in addition, unique N-linked glycosylation, transmembrane and cytosolic domains. A rabbit polyvalent antiserum was generated against residues 27-40 in the shared amino-terminal domain and a mouse polyvalent antiserum was generated against the full-length protein to study trafficking of individual UL37 proteins in human cells that transiently expressed gpUL37 or pUL37x1. Co-localization studies by confocal laser scanning microscopy detected trafficking of gpUL37 and pUL37x1 from the endoplasmic reticulum to the Golgi apparatus in permissive U373 cells and in human diploid fibroblasts (HFF). Trafficking of gpUL37 to the cellular plasma membrane was detected in unfixed HFF cells. FLAG-tagged gpUL37 trafficked similarly through the secretory apparatus to the plasma membrane. By using confocal microscopy and immunoblotting of fractionated cells, gpUL37 and pUL37x1 were found to co-localize with mitochondria in human cells. This unconventional dual trafficking pattern through the secretory apparatus and to mitochondria is novel for herpesvirus IE regulatory proteins.

Animals↗

Stoichiometry of the photosynthetic apparatus and phycobilisome structure of the cyanobacterium Plectonema boryanum UTEX 485 are regulated by both light and temperature.

The role of growth temperature and growth irradiance on the regulation of the stoichiometry and function of the photosynthetic apparatus was examined in the cyanobacterium Plectonema boryanum UTEX 485 by comparing mid-log phase cultures grown at either 29 degrees C/150 micromol m(-2) s(-1), 29 degrees C/750 micromol m(-2) s(-1), 15 degrees C/150 micromol m(-2) s(-1), or 15 degrees C/10 micromol m(-2) s(-1). Cultures grown at 29 degrees C/750 micromol m(-2) s(-1) were structurally and functionally similar to those grown at 15 degrees C/150 micromol m(-2) s(-1), whereas cultures grown at 29 degrees C/150 micromol m(-2) s(-1) were structurally and functionally similar to those grown at 15 degrees C/10 micromol m(-2) s(-1). The stoichiometry of specific components of the photosynthetic apparatus, such as the ratio of photosystem (PS) I to PSII, phycobilisome size and the relative abundance of the cytochrome b(6)/f complex, the plastoquinone pool size, and the NAD(P)H dehydrogenase complex were regulated by both growth temperature and growth irradiance in a similar manner. This indicates that temperature and irradiance may share a common sensing/signaling pathway to regulate the stoichiometry and function of the photosynthetic apparatus in P. boryanum. In contrast, the accumulation of neither the D1 polypeptide of PSII, the large subunit of Rubisco, nor the CF(1) alpha-subunit appeared to be regulated by the same mechanism. Measurements of P700 photooxidation in vivo in the presence and absence of inhibitors of photosynthetic electron transport coupled with immunoblots of the NAD(P)H dehydrogenase complex in cells grown at either 29 degrees C/750 micromol m(-2) s(-1) or 15 degrees C/150 micromol m(-2) s(-1) are consistent with an increased flow of respiratory electrons into the photosynthetic intersystem electron transport chain maintaining P700 in a reduced state relative to cells grown at either 29 degrees C/150 micromol m(-2) s(-1) or 15 degrees C/10 micromol m(-2) s(-1). These results are discussed in terms of acclimation to excitation pressure imposed by either low growth temperature or high growth irradiance.

Acclimatization↗

Involvement of the Golgi Apparatus in the Synthesis and Secretion of Hydroxyproline-rich Cell Wall Glycoproteins.

Pulse labeling of carrot root phloem parenchyma (Daucus carota L. cv. Nantes) tissue with (14)C-proline followed by fractionation of the cytoplasmic organelles on sucrose gradients was used to determine the identity of the membranous organelles involved in the secretion of the hydroxyproline-rich glycoproteins of the cell wall. Identification of the organelles was done through electron-microscopical observations and through the localization of marker enzymes on the sucrose gradients. Enrichment of the organelles involved in secretion was determined by measuring the percentage of the incorporated radioactivity present as (14)C-hydroxyproline. The Golgi apparatus (dictyosome) was found to be a major site of glycoprotein transport. This identification was based on the observed enrichment of dictyosomes paralleling the purification of newly synthesized cell-wall glycoproteins. A marker enzyme for the Golgi apparatus, inosinediphosphatase, banded with the newly synthesized cell wall glycoproteins on sequential isopycnic and rate zonal sucrose gradients. Marker enzymes for the endoplasmic reticulum and the plasma membrane were clearly separated from the dictyosome-rich fraction. UDP-arabinose arabinosyl transferase, an enzyme involved in the glycosylation of the peptide moiety of this glycoprotein, also banded with the dictyosomes on both kinds of gradients. The results suggest an important role of the Golgi apparatus in the biosynthesis and the secretion of the cell wall glycoproteins of higher plants.

Journal Article↗

Photosynthetic apparatus of pea thylakoid membranes : response to growth light intensity.

We investigated the effect of growth light intensity on the photosynthetic apparatus of pea (Pisum sativum) thylakoid membranes. Plants were grown either in a growth chamber at light intensities that ranged from 8 to 1050 microeinsteins per square meter per second, or outside under natural sunlight. In thylakoid membranes we determined: the amounts of active and inactive photosystem II, photosystem I, cytochrome b/f, and high potential cytochrome b(559), the rate of uncoupled electron transport, and the ratio of chlorophyll a to b. In leaves we determined: the amounts of the photosynthetic components per leaf area, the fresh weight per leaf area, the rate of electron transport, and the light compensation point. To minimize factors other than growth light intensity that may alter the photosynthetic apparatus, we focused on peas grown above the light compensation point (20-40 microeinsteins per square meter per second), and harvested only the unshaded leaves at the top of the plant. The maximum difference in the concentrations of the photosynthetic components was about 30% in thylakoids isolated from plants grown over a 10-fold range in light intensity, 100 to 1050 microeinsteins per square meter per second. Plants grown under natural sunlight were virtually indistinguishable from plants grown in growth chambers at the higher light intensities. On a leaf area basis, over the same growth light regime, the maximum difference in the concentration of the photosynthetic components was also about 30%. For peas grown at 1050 microeinsteins per square meter per second we found the concentrations of active photosystem II, photosystem I, and cytochrome b/f were about 2.1 millimoles per mol chlorophyll. There were an additional 20 to 33% of photosystem II complexes that were inactive. Over 90% of the heme-containing cytochrome f detected in the thylakoid membranes was active in linear electron transport. Based on these data, we do not find convincing evidence that the stoichiometries of the electron transport components in the thylakoid membrane, the size of the light-harvesting system serving the reaction centers, or the concentration of the photosynthetic components per leaf area, are regulated in response to different growth light intensities. The concept that emerges from this work is of a relatively fixed photosynthetic apparatus in thylakoid membranes of peas grown above the light compensation point.

Journal Article↗

Developmental Regulation of the Plastid Protein Import Apparatus.

Plastid development involves the programmed accumulation of proteins. Most plastid proteins are synthesized in the cytosol and imported into the organelle by an envelope-based protein import apparatus. Previous studies have shown that developmental rates of protein accumulation correspond to mRNA levels. Here, we examined the relationship between plastid development and the activity of the protein import apparatus. Developing plastids, primarily from wheat leaves, were analyzed for their protein import capability in vitro. Import capability, initially high in proplastids, declined as much as 20-fold as plastid development approached either the mature etioplast or the mature chloroplast. The observed decline was not due to senescence, nonspecific inhibitors, or protein turnover. Furthermore, the import capability of mature etioplasts, initially very low, was transiently reactivated during light-mediated redifferentiation into chloroplasts. These results suggest that plant cells regulate the import apparatus in concert with the protein demands of the developing plastids.

Journal Article↗

A new apparatus for polarized X-ray absorption fine structure using a grazing-incidence fluorescence excitation.

A new apparatus is described for surface-sensitive X-ray absorption fine structure (XAFS) at the 27-pole wiggler beamline BL13B at the Photon Factory. The apparatus is designed for polarized XAFS experiments at low temperature. A closed-cycle helium cryostat is rotated around the omega axis of a vertically mounted high-precision goniometer. The apparatus can be used for XAFS and X-ray standing-wave experiments, sharing a 19-element pure-Ge detector array as a fluorescence detector. The available temperature range is 15-300 K. The performance is demonstrated by in-plane polarized Cu K-EXAFS for a YBa2Cu307 thin film (approximately 1000 A) epitaxically grown on an SrTiO3 single crystal. An in-situ study of amorphous selenium under optical excitation revealed that neutral defect pairs (C3(0)-C3(0)) are formed by photoexcitation of lone pairs. This indicates that grazing-incidence fluorescence excitation is advantageous for 'pump-and-probe' experiments which can minimize the mismatch in the probing depth between X-ray and optical excitation.

Journal Article↗

An apparatus for characterization and control of isolated muscle.

An apparatus for characterization and control of muscle tissue is presented. The apparatus is capable of providing generalized mechanical boundary conditions to muscle tissue, as well as implementing real-time feedback control via electrical stimulation. The system is intended to serve as an experimental platform for implementing a wide variety of muscle control and identification studies that will serve as fundamental investigations of muscle mechanics, energetics, functional electrical stimulation, and fatigue. In one illustration of the capabilities of the apparatus, pilot experimental results of muscle workloops against a finite-admittance passive load are presented, illustrating how richer boundary conditions may reveal interesting muscle behavior.

Animals↗

Doctor Dee's ether apparatus.

In the early 1940s, Dr M. H. Armstrong-Davison devised an apparatus for inhalational anaesthesia. The apparatus was essentially a combination of Water's to-and-fro soda-lime canister and Hewitt's ether inhaler. The description of the apparatus is followed by a brief historical note on Dr Armstrong-Davison.

Anesthesia, Inhalation↗

A new apparatus for continuous cultivation of bacterial plaque on solid surfaces and human dental enamel.

A new apparatus for the continuous cultivation of mono and mixed bacterial plaque on solid surfaces is described. The features are: easy preparation and handling; freedom from technical problems and microbial contamination; self-sufficient for periods of up to 56 d; 12 samples are taken simultaneously; programmable supply inlet. Experiments were performed with Streptococcus mutans C 67-1 for mono bacterial inoculation and in combination with Veillonella alcalescens V-1 for mixed bacterial inoculations. The results showed that the controlled conditions and versatility of the apparatus make possible the study of plaque-development and lesion production on a time-dependent basis. It is concluded that the apparatus is suitable for a wide range of dental and non-dental applications.

Bacteria↗

Surface decontamination of beef inoculated with Salmonella Typhimurium DT104 or Escherichia coli O157:H7 using dry air in a novel heat treatment apparatus.

AIMS: To determine the effectiveness of a novel dry air decontamination apparatus in the deactivation of Salmonella serotype Typhimurium DT104 or Escherichia coli O157:H7 on beef surfaces. METHODS AND RESULTS: A laboratory scale dry air decontamination apparatus, capable of producing repeatable and known heating time-temperature cycles on food surfaces was used in decontamination trials. Beef samples were surface inoculated with 7-8 log10CFU cm(-2) of S. Typhimurium DT104 or E. coli O157:H7 and heated at 60, 75, 90 and 100 degrees C using fast and slow heating rates and subsequently held at these temperatures for up to 600 s. A substantial reduction in pathogen numbers was achieved at higher temperatures (90 and 100 degrees C, 4.18-6.06 log10CFU cm(-2)) using both heating rates, but cell survival at these temperatures was also observed. At the lower temperatures, deactivation was small at 60 degrees C in particular it was less than one log unit after 3 min heating. No significant differences were observed when total reductions in pathogen counts were compared for all the temperature/heat up time combinations tested. During slow heating at 90 degrees C, and both heating rates at 100 degrees C, the pattern of deactivation of S. Typhimurium DT104 or E. coli O157:H7 was triphasic. CONCLUSIONS: This study has shown that heating meat surfaces with dry air can achieve substantial reductions in S. Typhimurium DT104 or E. coli O157:H7. As surface decontamination of beef surfaces with dry air had a negative effect on beef colour and appearance, such a decontamination apparatus would be unsuitable for producing meat for retail sale but it could be used to produce safer meat for use in the catering trade. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides researchers and food processors with data on the dynamic changes in S. Typhimurium DT104 and E. coli O157:H7 counts on intact beef surfaces during heating with dry air under realistic (time-varying) temperature conditions.

Air↗

The type IV secretion apparatus protein VirB6 of Agrobacterium tumefaciens localizes to a cell pole.

Agrobacterium tumefaciens VirB proteins assemble a type IV secretion apparatus for the transfer of DNA and proteins to plant cells. To study the role of the VirB6 protein in the assembly and function of the type IV apparatus, we determined its subcellular location by immunofluorescence microscopy. In wild-type bacteria VirB6 localized to the cell poles but in the absence of the tumour-inducing plasmid it localized to random sites on the cell membranes. Five of the 11 VirB proteins, VirB7-VirB11, are required for the polar localization of VirB6. We identified two regions of VirB6, a conserved tryptophan residue at position 197 and the extreme C-terminus, that are essential for its polar localization. Topology determination by PhoA fusion analysis placed both regions in the cell cytoplasm. Alteration of tryptophan 197 or the deletion of the extreme C-terminus led to the mislocalization of the mutant protein. The mutations abolished the DNA transfer function of the protein as well. The C-terminus of VirB6, in silico, can form an amphipathic helix that may encode a protein-protein interaction domain essential for targeting the protein to a cell pole. We previously reported that another DNA transfer protein, VirD4, localizes to a cell pole. To determine whether VirB6 and VirD4 localize to the same pole, we performed colocalization experiments. Both proteins localized to the same pole indicating that VirB6 and VirD4 are in close proximity and VirB6 is probably a component of the transport apparatus.

Agrobacterium tumefaciens↗

A secreted anti-activator, OspD1, and its chaperone, Spa15, are involved in the control of transcription by the type III secretion apparatus activity in Shigella flexneri.

Bacteria of Shigella spp. are responsible for shigellosis in humans and use a type III secretion (TTS) system to enter epithelial cells and trigger apoptosis in macrophages. Transit of translocator and effector proteins through the TTS apparatus is activated upon contact of bacteria with host cells. Transcription of approximately 15 genes encoding effectors is regulated by the TTS apparatus activity and controlled by MxiE, an AraC family activator, and its coactivator IpgC, the chaperone of IpaB and IpaC translocators. Using a genetic screen, we identified ospD1 as a gene whose product negatively controls expression of genes regulated by secretion activity. OspD1 associates with the chaperone Spa15 and the activator MxiE and acts as an anti-activator until it is secreted. The mechanism regulating transcription in response to secretion activity involves an activator (MxiE), an anti-activator (OspD1), a co-anti-activator (Spa15), a coactivator (IpgC) and two anti-coactivators (IpaB and IpaC) whose alternative and mutually exclusive interactions are controlled by the duration of the TTS apparatus activity.

Bacterial Proteins↗

Elimination of apparatus dead space--a simple method for improving CO2 removal without increasing airway pressure.

During mechanical ventilation the apparatus dead space can be eliminated by insufflating through one lumen of a tracheal double-lumen tube and allowing expiration through the other. In six intravenously anesthetized pigs, this technique resulted in an 18% (1 kPa) decrease in PaCO2 compared with insufflating through both lumens (32 ml rebreathing volume). Oxygenation, airway pressures, and tidal volumes were unchanged. Flushing the trachea with fresh gas during the expiratory phase did not improve the efficiency of ventilation. It is concluded that elimination of apparatus dead space improves CO2 removal without increasing airway pressures and tidal volumes, and it is suggested that minimization of apparatus dead space should be tried before more advanced ventilatory modes are considered.

Animals↗

L-tyrosine, L-dopa, and tyrosinase as positive regulators of the subcellular apparatus of melanogenesis in Bomirski Ab amelanotic melanoma cells.

In cultured cells of the Bomirski Ab amelanotic hamster melanoma line, the substrates of tyrosinase, L-tyrosine, and L-DOPA induce the melanogenic pathway. In this report, we demonstrate that these substrates regulate the subcellular apparatus involved in their own metabolism and that this regulation is under the dynamic control of one of the components of this apparatus, tyrosinase, via tyrosine hydroxylase activity. Culturing cells with nontoxic but melanogenically inhibitory levels of phenylthiourea (PTU; 100 microM) strongly inhibits induction of both the tyrosine hydroxylase and DOPA oxidase activities of tyrosinase by L-tyrosine (200 microM) but has no effect on the induction of either activity by L-DOPA (50 microM). De novo synthesis of premelanosomes precedes the onset of tyrosine-induced melanogenesis. Thereafter, increases in the population of melanosomes (likewise inhibited by PTU) correlate positively with increases in tyrosinase activity induced by L-tyrosine. Melanogenesis induced by L-DOPA in the absence of L-tyrosine is rate-limited not by tyrosinase but by inadequate melanosome synthesis. Our findings indicate that in Bomirski Ab amelanotic hamster melanoma cells the synthesis of the subcellular apparatus of melanogenesis is initiated by L-tyrosine and is regulated further by tyrosinase and L-DOPA, which serves as a second messenger subsequent to tyrosine hydroxylase activity.

Animals↗

Magnetic resonance imaging of the palmar aspect of the equine podotrochlear apparatus: normal appearance.

The purpose of this study was to describe the normal magnetic resonance (MR) imaging characteristics of the palmar structures of the equine podotrochlear apparatus by means of retrospective evaluation of MR imaging studies of 16 cadaver limbs. The articular aspect of the distal sesamoid bone was not evaluated in this study. Equine digits were imaged with a human knee radiofrequency coil in a 1.5 T magnetic field, using spin echo (SE) T1-weighted, turbo spin echo proton density (TSE PD)-weighted with and without fat saturation (FS), and FS TSE T2-weighted sequences. The limbs were dissected after imaging to validate the absence of gross abnormalities of the flexor aspect of the distal sesamoid bone, of the deep digital flexor tendon, and the distal impar sesamoidean ligament. Seven deep digital flexor tendons were subjected to histologic examination to exclude any microscopic tendon pathology. The anatomic structures of the podotrochlear apparatus were easily identified on MR images. Compact bone of the flexor cortex of the distal sesamoid bone had low intensity signal on all sequences. In 11 digits an increased signal was seen within the thickness of the sagittal eminence of the flexor cortex in SE T1-weighted images and in TSE PD-weighted images without FS. Trabecular bone had a granular appearance and high signal in SE T1-weighted sequences and TSE images without FS. The deep digital flexor tendon had low signal on FS T2-weighted images, while on short echo time sequences (T1- and PD-weighted sequences), the tendon signal varied depending on the relative orientation between its fibers and the static magnetic field. Seven tendons had stippled appearance due to small intratendonous foci of slightly increased signal on transverse T1-weighted images. MR imaging provides a thorough evaluation of the anatomical structure of the podotrochlear apparatus: A good knowledge of the MR imaging appearance and anatomy and an awareness of potential pitfalls will improve diagnostic specificity for the detection of pathologic changes.

Animals↗

An apparatus for studying rapid electrophysiological responses to light demonstrated on Arabidopsis leaves.

An apparatus for making high-resolution measurements of electrophysiological changes induced by light in plant cells was constructed. Its main components were a xenon arc lamp, an electronic shutter, a liquid light-guide, a computer equipped with an analog-to-digital converter and a computer program that controlled the shutter and data acquisition. The apparatus was used to examine transient changes in membrane potential (Vm) that occur upon illumination in Arabidopsis leaves. Light-on induced a transient hyperpolarization of 4 mV after a lag time of 0.53 s. It was followed by a much larger transient depolarization that peaked 31 s after light-on. The Vm returned to near its original value after approximately 3 min. The early changes in Vm have been proposed to result from effects of photosynthetically produced ATP on the activities of H(+)-ATPases and K+ channels at the plasma membrane. The kinetics of the initial hyperpolarization were found to be reasonably consistent with such a mechanism. It is expected that the apparatus described here will be useful in future investigations of this and other electrophysiological responses to light.

Adenosine Triphosphate↗