High -resolution scanning electron-microscopic studies on the three-dimensional structure of mitochondria and sarcoplasmic reticulum in the different twitch muscle fibers of the frog

Summary The three-dimensional structure of the mitochondria and sarcoplasmic reticulum (SR) in the three types of twitch fibers, i.e., the red, white and intermediate skeletal muscle fibers, of the vastus lateralis muscle of the Japanese meadow frog (Rana nigromaculata nigromaculata Hallowell) was examined by high resolution scanning electron microscopy, after removal of the cytoplasmic matrices.The small red fibers have numerous mitochondrial columns of large diameter, while the large white fibers have a small number of mitochondrial columns of small diameter. In the medium-size intermediate fibers, the number and diameter of the mitochondrial columns are intermediate between those of the red and white fibers.In all three types of fibers, the terminal cisternae and transverse tubules form triads at the level of each Z-line. The thick terminal cisternae continue into much thinner flat intermediate cisternae, through a transitional part where a row of tiny indentations can be observed. Numerous slender longitudinal tubules originating from the intermediate cisternae, extend longitudinally or obliquely and form elongated oval networks of various sizes in front of the A-band, then fuse to form the H-band collar (fenestrated collar) around the myofibrils. On the surface of the H-band collar, small fenestrations as well as tiny hollows are seen. The three-dimensional structure of SR is basically the same in all three muscle fiber-types. However, the SR is sparse on the surface of mitochondria, so the mitochondria-rich red fiber has a smaller total volume of SR than the mitochondria-poor white fiber. The volume of SR of the intermediate fiber is intermediate between other the two.     

Scanning electron-microscopic studies on the three-dimensional structure of sarcoplasmic reticulum in the mammalian red,white and intermediate muscle fibers

Summary The three-dimensional structure of the sarcoplasmic reticulum (SR) in the red, white and intermediate striated muscle fibers of the extensor digitorum longus muscle of the rat was examined under a field-emission type scanning electron microscope after removal of cytoplasmic matrices by the osmium-DMSO-osmium procedure.In all three types of fibers, the terminal cisternae and transverse tubules form triads at the level of the A-I junction. Numerous slender sarcotubules, originating from the A-band side terminal cisternae, extend obliquely or longitudinally and form oval or irregular shaped networks of various sizes in front of the A-band, then become continuous with the tiny mesh (fenestrated collar) in front of the H-band. The A-and H-band SR appears as a single sheet of anastomotic tubules. Numerous sarcotubules, originating from the I-band side terminal cisternae, extend in threedimensional directions and form a multilayered network over the I-band and Z-line regions. At the I-band level, paired transversely oriented mitochondria partly embrace the myofibril. The I-band SR network is poorly developed in the narrow space between the paired mitochondria, but is well developed in places devoid of these mitochondria.The three-dimensional structure of the SR is basically the same in all three muscle fiber-types. However, the SR is sparse on the surface of mitochondria, so the mitochondria-rich red fiber has a much smaller total volume of SR than the mitochondria-poor white fiber. Moreover, the volume of SR of the intermediate fiber is intermediate between the two.     

A correlative transmission and scanning electron microscopic study of the pigeon myocardial cell

Summary A comparative study of the pigeon ventricular myocardial cell has been performed by transmission electron microscopy (TEM) and by scanning electron microscopy (SEM). Three-dimensional access to the cell interior was obtained by cryo-fracturing paraffin-embedded tissue immersed in liquid nitrogen. The TEM studies revealed parallelly arranged myofibrils separated by rows of mitochondria. The sarcoplasmic reticulum is represented by a well-developed network of tubules which, at the Z- and H-band level of the sarcomere, expands to form belt-like cisternae. The cisternae at the Z-band level lie in close proximity to both myofilaments and mitochondria. Transverse tubules are absent and thus only peripheral couplings are present.SEM observations of the fractured tissue revealed the spatial relationship between the different cell organelles, the most important of these being the parallel myofibrils and the mitochondria. The conspicuous ridges transversing the myofibril at the Z-band level consist mainly of expanded Z-bands, but overlying SR-tubules also contribute to these ridges. Traces of the SR can sometimes be seen covering the myofibrils. The close proximity between the SR and the mitochondria was also confirmed in the SEM.Preparation and examination of SEM prepared tissue in the TEM confirmed that no essential damage or reorganization of cell organelles had taken place during the SEM procedure. On the other hand some shrinkage of the tissue, which was probably caused by critical point drying, was noticed.     

Fine structure and differentiation of Ascidian muscle. I. Differentiated caudal musculature of Distaplia occidentalis tadpoles

The structure of the caudal muscle in the tadpole larva of the compound ascidian Distaplia occidentalis has been investigated with light and electron microscopy. The two muscle bands are composed of about 1500 flattened cells arranged in longitudinal rows between the epidermis and the notochord. The muscle cells are mononucleate and contain numerous mitochondria, a small Golgi apparatus, lysosomes, proteid-yolk inclusions, and large amounts of glycogen. The myofibrils and sarcoplasmic reticulum are confined to the peripheral sarcoplasm. Myofibrils are discrete along most of their length but branch near the tapered ends of the muscle cell, producing a Felderstruktur. The myofibrils originate and terminate at specialized intercellular junctional complexes. These myomuscular junctions are normal to the primary axes of the myofibrils and resemble the intercalated disks of vertebrate cardiac muscle. The myofibrils insert at the myomuscular junction near the level of a Z-line. Thin filaments (presumably actin) extend from the terminal Z-line and make contact with the sarcolemma. These thin filaments frequently appear to be continuous with filaments in the extracellular junctional space, but other evidence suggests that the extracellular filaments are not myofilaments. A T-system is absent, but numerous peripheral couplings between the sarcolemma and cisternae of the sarcoplasmic reticulum (SR) are present on all cell surfaces. Cisternae coupled to the sarcolemma are continuous with transverse components of SR which encircle the myofibrils at each I-band and H-band. The transverse component over the I-band consists of anastomosing tubules applied as a single layer to the surface of the myofibril. The transverse component over the H-band is also composed of anastomosing tubules, but the myofibrils are invested by a double or triple layer. Two or three tubules of sarcoplasmic reticulum interconnect consecutive transverse components. Each muscle band is surrounded by a thin external lamina. The external lamina does not parallel the irregular cell contours nor does it penetrate the extracellular space between cells. In contracted muscle, the sarcolemmata at the epidermal and notochordal boundaries indent to the level of each Z-line, and peripheral couplings are located at the base of the indentations. The external lamina and basal lamina of the epidermis are displaced toward the indentations. The location, function, and neuromuscular junctions of larval ascidian caudal muscle are similar to vertebrate somatic striated muscle. Other attributes, including the mononucleate condition, transverse myomuscular junctions, prolific gap junctions, active Golgi apparatus, and incomplete nervous innervation are characteristic of vertebrate cardiac muscle cells.     

The membrane systems of the cardiac muscle cell of Euchaeta norvegica Boeck (Crustacea,Copepoda)

Summary The membrane systems of the cardiac muscle cell of the copepod Euchaeta norvegica Boeck are described. The heart wall, which is between 0.12 and 1.36 m thick, consists of an epicardium and a single layer of muscle cells. Invaginations of the sarcolemma forming transverse tubules have been found at all levels of the sarcomere with the exception of the H-band level. The longitudinal tubules of the same system are closely associated with the sarcoplasmic reticulum to form interior couplings at the A-I level of the sarcomere. Triadic couplings at the Z band level were not seen in E. norvegica, but peripheral couplings were demonstrated. Nexuses were found in the intercalated discs.     

Eubacterium aggreganssp. nov., a New Homoacetogenic Bacterium from Olive Mill Wastewater Treatment Digestor

A strictly anaerobic, homoacetogenic, Gram-positive, non spore-forming bacterium, designated strain SR12^T(T=type strain), was isolated from an anaerobic methanogenic digestor fed with olive mill wastewater. Yeast extract was required for growth but could also be used as sole carbon and energy source. Strain SR12^Tutilized a few carbohydrates (glucose, fructose and sucrose), organic compounds (lactate, crotonate, formate and betaine), alcohols (methanol), the methoxyl group of some methoxylated aromatic compounds, and H₂+CO₂. The end-products of carbohydrate fermentation were acetate, formate, butyrate, H₂and CO₂. End-products from lactate and methoxylated aromatic compounds were acetate and butyrate. Strain SR12^Twas non-motile, formed aggregates, had a G+C content of 55 mol % and grew optimally at 35°C and pH 7.2 on a medium containing glucose. Phylogenetically, strain SR12^Twas related toEubacterium barkeri, E. callanderi, andE. limosumwithE. barkerias the closest relative (similarity of 98%) with which it bears little phenotypic similarity or DNA homology (60%). On the basis of its phenotypic, genotypic, and phylogenetic characteristics, we propose to designate strain SR12^TasEubacterium aggreganssp. nov. The type strain is SR12^T(=DSM 12183).     

High resolution scanning electron-microscopic study on the three-dimensional structure of the sarcoplasmic reticulum in the slow (tonic) muscle fibers of the frog,Rana nigromaculata

Summary The three-dimensional structure of the sarcoplasmic reticulum (SR) in the slow (tonic) fibers of the reclus abdominis muscle of the Japanese meadow frog (Rana nigromaculata nigromaculata Hallowell) was examined by high resolution scanning electron microscopy, after removal of the cytoplasmic matrices by the osmium-DMSO-osmium procedure. The SR forms a repetitive network throughout these fibers. At the level of the Z-line, a slender transverse tubule (T-tubule) runs transversely to the longitudinal axis of the myofibril. Small, spherical or ovoid terminal cisternae couple laterally with the T-tubule at intervals of 0.4–1.0 m, and form a terminal cisterna-T-tubule complex on whose surface tiny indentations are occasionally seen. Each terminal cisterna gives rise to a few sarcotubules that run in various directions, divide frequently and form circular or oval meshes of diverse sizes in front of the A- and I-bands. The sarcotubules usually form small meshes in the middle of the A-band, but occasionally fuse and form a poorly developed H-band (fenestrated) collar.     

Ultrastructure of the myocardial cell and its membrane systems in the adult fly Calliphora erythrocephala (Insecta: Diptera)

Summary Calliphora erythrocephala has cross-striated cardiac muscle cells with A, I and Z-bands. The diameters of the myosin and actin filaments are 200–250 Å and 85 Å respectively and the length of the myosin filaments (A-band) is approximately 1.5 . Usually 8–10 actin filaments surround each myosin filament.The myocardial cells show a well-developed membrane system and interior couplings. A perforated sheet of SR envelopes the myofibrils at the A-band, dilates into flattened cisternae at both A-I band levels before it merges into a three-dimensional net-work between the actin filaments of the I-bands and between the dense bodies of the discontinuous Z-discs. The T-system consists of broad flattened tubules running between the myofibrils at the A-I band levels forming dyads with the SR-cisternae. Longitudinal connections between the transverse (T-) tubules often occur.It is suggested that this well-developed SR may be an adaptation to facilitate a rapid contraction/relaxation frequency by an effective Ca²⁺ uptake.     

Body temperature and thermoregulation ofAntheraea assama larvae

Activity patterns and body temperature (T _b ) and its maintenance were investigated in the 5th instar larvae ofAntheraea assama Westwood (Lepidoptera: Saturniidae). These larvae thermoregulate by finding and utilizing favourable microenvironments and by restricting their activities to suitable times of the day. Further convective cooling, facilitated by a raised posture also serves to ameliorate T_b. Regression analysis suggests that environmental temperature is the major determinant of T_b along with solar radiation (SR) and wind (W).     

Penetration of horseradish peroxidase into the terminal cisternae of frog skeletal muscle fibers and blockade of caffeine contracture by Ca++ depletion

Summary Horseradish peroxidase, an extracellular marker, was given intravenously to frogs, and 40 min later the sartorius muscles were removed. The isolated muscles were exposed for an additional hour to Ringer solution containing peroxidase, then fixed with glutaraldehyde. Peroxidase activity was found in the T tubules, in some of the terminal cisternae (TC) of the SR, and occasionally in the longitudinal tubules of the SR. In transverse sections, the structures containing tracer formed a pattern of approximately parallel columns reaching to the cell surface; the statistical distribution of their spacing was nearly the same as that of the interdistances between the current-sensitive spots on the Z-line which triggered localized contraction (Huxley and Taylor, 1958). The caffeine contracture of frog sartorius muscles remained unchanged in isotonic Ringer solutions which were Ca⁺⁺-free or contained Mn⁺⁺ or La⁺⁺⁺; however, contracture was blocked by prior exposure of the muscles to the same solutions made 2 × hypertonic with sucrose (known to produce swelling of T tubules and (TC). Since Mn⁺⁺ and La⁺⁺⁺ are known to depress Ca⁺⁺ influx, these results suggest that washout of Ca⁺⁺ from the TC, and penetration of La⁺⁺⁺ or Mn⁺⁺ into it, occur more rapidly due to the swelling of T tubules and TC associated with hypertonicity. It is concluded that at least some of the terminal cisternae are open to the interstitial fluid via the T tubules. Thus, depolarization of the T tubules could readly depolarize the cisternae and lead to Ca⁺⁺ influx into the myoplasm.Supported by grants from the Public Health Service (HE-11155, HE-05815, and HE-10384) and from the American Heart Assocation. The authors are indebted to Mrs. Jan Redick for expert technical assistance.     

Penetration of horseradish peroxidase into the terminal cisternae of frog skeletal muscle fibers and blockade of caffeine contracture by Ca++ depletion

Summary Horseradish peroxidase, an extracellular marker, was given intravenously to frogs, and 40 min later the sartorius muscles were removed. The isolated muscles were exposed for an additional hour to Ringer solution containing peroxidase, then fixed with glutaraldehyde. Peroxidase activity was found in the T tubules, in some of the terminal cisternae (TC) of the SR, and occasionally in the longitudinal tubules of the SR. In transverse sections, the structures containing tracer formed a pattern of approximately parallel columns reaching to the cell surface; the statistical distribution of their spacing was nearly the same as that of the interdistances between the current-sensitive spots on the Z-line which triggered localized contraction (Huxley and Taylor, 1958). The caffeine contracture of frog sartorius muscles remained unchanged in isotonic Ringer solutions which were Ca⁺⁺-free or contained Mn⁺⁺ or La⁺⁺⁺; however, contracture was blocked by prior exposure of the muscles to the same solutions made 2 × hypertonic with sucrose (known to produce swelling of T tubules and (TC). Since Mn⁺⁺ and La⁺⁺⁺ are known to depress Ca⁺⁺ influx, these results suggest that washout of Ca⁺⁺ from the TC, and penetration of La⁺⁺⁺ or Mn⁺⁺ into it, occur more rapidly due to the swelling of T tubules and TC associated with hypertonicity. It is concluded that at least some of the terminal cisternae are open to the interstitial fluid via the T tubules. Thus, depolarization of the T tubules could readly depolarize the cisternae and lead to Ca⁺⁺ influx into the myoplasm.Supported by grants from the Public Health Service (HE-11155, HE-05815, and HE-10384) and from the American Heart Assocation. The authors are indebted to Mrs. Jan Redick for expert technical assistance.     

<Emphasis Type="Italic">Desulfurispirillum alkaliphilum</Emphasis> gen. nov. sp. nov., a novel obligately anaerobic sulfur- and dissimilatory nitrate-reducing bacterium from a full-scale sulfide-removing bioreactor

Strain SR 1^T was isolated under anaerobic conditions using elemental sulfur as electron acceptor and acetate as carbon and energy source from the Thiopaq bioreactor in Eerbeek (The Netherlands), which is removing H₂S from biogas by oxidation to elemental sulfur under oxygen-limiting and moderately haloalkaline conditions. The bacterium is obligately anaerobic, using elemental sulfur, nitrate and fumarate as electron acceptors. Elemental sulfur is reduced to sulfide through intermediate polysulfide, while nitrate is dissimilatory reduced to ammonium. Furthermore, in the presence of nitrate, strain SR 1^T was able to oxidize limited amounts of sulfide to elemental sulfur during anaerobic growth with acetate. The new isolate is mesophilic and belongs to moderate haloalkaliphiles, with a pH range for growth (on acetate and nitrate) from 7.5 to 10.25 (optimum 9.0), and a salt range from 0.1 to 2.5 M Na⁺ (optimum 0.4 M). According to phylogenetic analysis, SR 1^T is a member of a deep bacterial lineage, distantly related to Chrysiogenes arsenatis (Macy et al. 1996). On the basis of the phenotypic and genetic data, the novel isolate is placed into a new genus and species, Desulfurispirillum alkaliphilum (type strain SR^T = DSM 18275 = UNIQEM U250). Nucleotide sequence accession number: the GenBank/EMBL accession number of the 16S rRNA gene sequence of strain SR 1^T is DQ666683.     

The ultrastructure of the striated muscle of the tail of Cercaria chackai Nadakal et al., 1969

The electron microscopic study of the tail of Cercaria chackai reveals that it contains four sets of striated muscle bundles located central to the nonstriated circular and longitudinal muscles. The striated muscle consists of longitudinally oriented lamellar myofibres. Each myofibre contains a single "U" shaped myofibril. The banding pattern is analogous to that of vertebrate striated muscle. The sarcolemma is a simple surface membrane. There are no transverse tubular extensions of sarcolemma. The sarcoplasmic reticulum (SR) is very well developed with cisternae, tubules, and vesicles. SR cisternae form dyadic couplings with the sarcolemma. There is a set of flattened tubules of SR origin traversing the myofibril exactly at the Z region. These tubules are unique to the striated muscle of the cercarian tail and may have functional significance. A diagrammatic reconstruction of the myofibre is presented.     

The tubular network of the Golgi apparatus

 Golgi apparatus of both plant and animal cells are characterized by an extensive system of approximately 30 nm diameter peripheral tubules. The total surface area of the tubules and associated fenestrae is thought to be approximately equivalent to that of the flattened portions of cisternae. The tubules may extend for considerable distances from the stacks. The tubules are continuous with the peripheral edges of the stacked cisternae, but the way they interconnect differs across the stack. In plant cells, for example, tubules associated with the near-cis and mid cisternae often begin to anastomose close to the peripheral edges of the stacked cisternae, whereas the tubules of the trans cisternae are less likely to anastomose and are more likely to be directly continuous with the peripheral edges of the stacked cisternae. Additionally, the tubules may blend gradually into fenestrae that surround some of the stack cisternae. Because of the large surface area occupied by tubules and fenestrae, it is reasonable to suppose that these components of the Golgi apparatus play a significant role in Golgi apparatus function. Tubules clearly interconnect closely adjacent stacks of the Golgi apparatus and may represent a communication channel to synchronize stack function within the cell. A feasible hypothesis is that tubules may be a potentially static component of the Golgi apparatus in contrast to the stacked cisternal plates which may turn over continuously. The coated buds associated with tubules may represent the means whereby adjacent Golgi apparatus stacks exchange carbohydrate-processing enzymes or where resident Golgi apparatus proteins are introduced into and out of the stack during membrane flow differentiation. The limited gradation of tubules from cis to medial to trans offers additional possibilities for functional specialization of Golgi apparatus in keeping with the hypothesis that tubules are repositories of resident Golgi apparatus proteins protected from turnover during the flow differentiation of the flattened saccules of the Golgi apparatus stack. Accepted: 3 November 1997     

Coordinated development of myofibrils, sarcoplasmic reticulum and transverse tubules in normal and dysgenic mouse skeletal muscle, in vivo and in vitro

We studied the development of transverse (T)-tubules and sarcoplasmic reticulum (SR) in relationship to myofibrillogenesis in normal and dysgenic (mdg/mdg) mouse skeletal muscle by immunofluorescent labeling of specific membrane and myofibrillar proteins. At E16 the development of the myofibrils and membranes in dysgenic and normal diaphragm was indistinguishable, including well developed myofibrils, a delicate network of T-tubules, and a prominent SR which was not yet cross-striated. In diaphragms of E18 dysgenic mice, both the number and size of muscle fibers and myofibrillar organization were deficient in comparison to normal diaphragms, as previously reported. T-tubule labeling was abnormal, showing only scattered tubules and fragments. However, many muscle fibers displayed cross striation of sarcomeric proteins and SR comparable to normal muscle. In cultured myotubes, cross-striated organization of sarcomeric proteins proceeded essentially in two stages: first around the Z-line and later in the A-band. Sarcomeric organization of the SR coincided with the first stage, while the appearance of T-tubules in the mature transverse orientation occurred infrequently, only after A-band maturation. In culture, myofibrillar and membrane organization was equivalent in normal and dysgenic muscle at the earlier stage of development, but half as many dysgenic myotubes reached the later stage as compared to normal. We conclude that the mdg mutation has little effect on the initial stage of membrane and myofibril development and that the deficiencies often seen at later stages result indirectly from the previously described absence of dihydropyridine receptor function in the mutant.     

Connections between dictyosomes,ER and GERL in cotyledons of mung bean (Vigna radiata L.)

Summary The connections and structural inter-relations of dictyosomes and endoplasmic reticulum (ER) in cotyledons of germinating mung beans were studied using thick (0.3 m) sections of aldehyde fixed, zinc iodide-osmium tetroxide (ZIO) impregnated tissue. The sections were examined by conventional (100 kV), rather than high voltage, transmission electron microscopy.Continuity of cisternal ER with tubular ER was confirmed and a direct connection of tubular ER totrans dictyosome cisternae was observed as were GERL networks associated withtrans dictyosome cisternae.Dictyosomes also gave rise to an extensive system of very fine tubules (10–20 nm diam) which have not been described previously in plant tissue. These tubules, which originated at thetrans dictyosome face, extended throughout the cytoplasm and were found connected to cisternal ER and tubular ER.The implications of these observations are discussed with regard to present ideas concerning endomembrane flow and protein sorting by the Golgi apparatus.     

Preparation and characterization of longitudinal tubules of sarcoplasmic reticulum from fast skeletal muscle

Sarcoplasmic reticulum (SR) serves a central role in calcium uptake and release, thereby regulating muscle relaxation and contraction, respectively. Recently, we have isolated fractions referable to longitudinal tubules (R2) and terminal cisternae (R4), the two major types of sarcoplasmic reticulum (A. Saito et al. (1984) J. Cell Biol. 99, 875-885). The terminal cisternae contain two types of membranes, the calcium pump membrane and the junctional face membrane. The terminal cisternae are filled with electron-opaque contents which serve as a Ca2+ reservoir. The longitudinal tubules consist mainly of the calcium pump membrane. In this study, we describe a new longitudinal tubule fraction (F2) and characterize it together with the R2 and R4 SR fractions. The calcium pump membrane of the longitudinal tubules is a highly specialized membrane consisting of about 90% calcium pump protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Extensive changes in morphology can be observed in the SR fractions referable to osmotic differences during the fixation conditions using either glutaraldehyde-tannic acid or osmium tetroxide fixatives. The changes include swelling or shrinkage and aggregation of the compartmental contents when the fixative contains calcium ions. The two types of SR have different osmotic permeability to the same medium, as indicated by differential swelling or shrinkage. Both longitudinal tubule and terminal cisternae vesicles of SR appear larger and are spherical vesicles when the glutaraldehyde-tannic acid fixative is isotonic as compared with the "standard" fixation method. We have previously reported that the ruthenium red-sensitive calcium release channels are localized to the terminal cisternae. The terminal cisternae as isolated are leaky to Ca2+ since these channels are in the "open state" (S. Fleischer et al. (1985) Proc. Natl. Acad. Sci USA 82, 7256-7259). Thus, the Ca2+, Mg2+-dependent ATPase (Ca2+ ATPase) rate is only slightly enhanced in the presence of a Ca2+ ionophore, which dissipates the Ca2+ gradient across the SR membrane. We now find that preincubation with ruthenium red restores the tight coupling of the Ca2+ ATPase activity to Ca2+ transport. That is to say, ATPase activity is reduced and the addition of ionophore stimulates the Ca2+ ATPase activity 4- to 7-fold. The Ca2+ ATPase activity in longitudinal tubules is already tightly coupled. It is minimal after a Ca2+ gradient has been generated, but can be stimulated 9- to 20-fold when the Ca2+ gradient is dissipated with ionophore. This finding suggests that the Ca2+ ATPase activity in SR is tightly coupled to Ca2+ transport in situ.     

Bending of z-lines by mechanical stimuli: An input signal for integrin dependent modulation of ion channels?

We studied which components of mechanical cell deformation are involved in “stretch modulated ion currents” (SMIC). Murine ventricular myocytes were attached to glass coverslips and deformed in x, y and z with a 16 μm thin glass stylus (S) of calibrated stiffness. Three-dimensional confocal microscopy characterized cell deformation (T-tubular membranes, mitochondria) and bending of S (indicative of the applied force). Axial (x-) displacement of S sheared the upper cell part versus the attached bottom, close to S, it changed sarcomere length and bent z-lines (“z-line displacement”). Vertical (z-press) or transversal (y-shear) displacement of S bulged cytoplasm and mitochondria transversally without detectable z-line displacement.Axial stiffness increased with the extent of stress (“stress stiffening”). Depolymerization of F-actin or block of integrin receptors reduced stiffness. SMIC served as a proxy readout of deformation-induced signaling. Axial deformation activated a non-selective cation conductance (G_ns) and deactivated an inwardly rectifying K⁺ conductance (G_K1), z-press or y-shear did not induce SMIC. Depolymerization of F-actin or block of integrin receptors reduced SMIC. SMIC did not depend on changes in sarcomere length but correlated with the extent of z-line bending. We discuss that both shear stress at the attached cell bottom and z-line bending could activate mechanosensors. Since SMIC was absent during deformations without z-line bending we postulate that z-line bending is a necessary component for SMIC signaling.     

Effect of non-thermal processing by High Hydrostatic Pressure on the survival of probiotic microorganisms: study on Bifidobacteria spp

High Hydrostatic Pressure (HP) processing has been suggested as an alternative method to improve textural attributes of dairy products. Since, the global market seeks improved functional foods, it is important to investigate whether HP processing can be applied to fermented dairy probiotic products. The inactivation kinetics of Bifidobacterium spp. in a model system of acid pH value under high pressure (100–400 MPa) combined with moderate temperature (20–35 °C) was investigated. Bifidobacterium spp. inactivation followed first order kinetics at all pressure–temperature combinations used. Pressure and temperature were found to act synergistically on the viability loss of the bacterium. The corresponding z_T and z_P values of inactivation were also estimated and, values of 41.5 °C and 93.5 MPa at reference pressure of 200 MPa and reference temperature of 25 °C were estimated, respectively. HP treatment of 200 MPa at 20–25 °C for 10–15 min, recommended for textural modification, is not detrimental to the viability of the studied probiotic culture and would be suitable for respective fermented probiotic products.     

Diploidization and chromosomal pairing affinities in the tetraploid wheats and their putative amphiploid progenitor

Summary The genomes of the diploid wheats Triticum boeoticum and T. urartu are closely related, giving 7II in the f₁ hybrid (T^bT^u) and 8.4 (0–14) II + 2.5 (0–7) IV in the derived amphiploid (T^bT^bT^uT^u). The genomes of the tetraploid wheats are also closely related, giving up to 7II at the polyhaploid level (AB) in the absence of the gene Ph but 14II at the tetraploid level (AABB) in the normal presence of Ph. If the amphiploid is the progenitor of the tetraploids, one or the other homoeologue (T^b or T^u) in each of the 7 homoeologous groups (the 7 potential IV) must have differentiated with respect to pairing affinity in order to account for 14II in the tetraploid. Consequently, in tetraploid X amphiploid hybrids (T^bT^uAB) carrying the Ph gene from the tetraploid, the seven differentiated chromosomes (B) would be expected to give 7I while, on the basis of their observed chiasma frequency, T^b, T^u and the less differentiated A would be expected to give 4.17I + 3.57II + 3.23III), assuming homoeologous pairing. The expected chromosomal configuration freqencies at MI (11.17I + 3.57II + 3.23III) closely fit the observed values (11.22I + 3.45II + 3.19III + 0.071IV) for such hybrids (X² = 0.0046; P>0.99). Thus diploidization of the boeoticum-urartu amphiploid clearly could account for the origin of the tetraploid wheats. Furthermore, T. aestivum X amphiploid hybrids (T^bT^uABD) with and without Ph indicated that B as well as A chomosomes tended to pair with their presumed T^bT^u homologues in the absence of Ph. Other tests showed that the tetraploid wheats could not plausibly have originated from any postulated Triticum-Sitopsis (TTSS) parental combinations with or without such chromosomal differentiation.