Rapid Contraction Phase Research Articles

Inflammation-Induced Lactate Leads to Rapid Loss of Hepatic Tissue-Resident NK Cells

The liver harbors two main innate lymphoid cell (ILC) populations: conventional NK (cNK) cells and tissue-resident NK (trNK) cells. Using the MCMV model of infection, we find that, in contrast to liver cNK cells, trNK cells initially undergo a contraction phase followed by a recovery phase to homeostatic levels. The contraction is MCMV independent because a similar phenotype is observed following poly(I:C)/CpG or α-GalCer injection. The rapid contraction phase is due to apoptosis, whereas the recovery phase occurs via proliferation in situ. Interestingly, trNK cell apoptosis is not mediated by fratricide and not induced by liver lymphocytes or inflammatory cytokines. Instead, we find that trNK cell apoptosis is the consequence of an increased sensitivity to lactic acid. Mechanistic analysis indicates that trNK cell sensitivity to lactate is linked to impaired mitochondrial function. These findings underscore the distinctive properties of the liver-resident NK cell compartment.

Inflammation-induced lactate leads to the rapid loss of hepatic tissue resident NK cells

Abstract The liver harbors two main innate lymphoid cell (ILC) populations, conventional NK (cNK) cells and tissue-resident NK (trNK) cells also called ILC1s. While the antiviral functions of cNK cells have been well established, trNK cell functions are only beginning to be uncovered. Using the MCMV model of infection, we found that in contrast to liver cNK cells, the trNK cell population initially undergoes a contraction phase prior to a recovery phase to homeostatic levels. The rapid contraction phase is due to apoptosis, while the recovery phase occurs via proliferation in situ. Liver trNK cell apoptosis is not mediated by fratricide, liver lymphocytes, or inflammatory cytokines. Instead, we found that liver trNK cell apoptosis is the consequence of an increased sensitivity to lactic acid. Interestingly, MCMV infection lead to an increase of lactate in the liver, which was supported by changes in lactate dehydrogenase expression. Mechanistic analysis indicates that trNK cell sensitivity to lactate is linked to impaired mitochondrial functions. These findings underscore the unique properties of the liver resident NK cell compartment.

Pelvic muscle rehabilitation: a standardized protocol for pelvic floor dysfunction.

Introduction. Pelvic floor dysfunction syndromes present with voiding, sexual, and anorectal disturbances, which may be associated with one another, resulting in complex presentation. Thus, an integrated diagnosis and management approach may be required. Pelvic muscle rehabilitation (PMR) is a noninvasive modality involving cognitive reeducation, modification, and retraining of the pelvic floor and associated musculature. We describe our standardized PMR protocol for the management of pelvic floor dysfunction syndromes. Pelvic Muscle Rehabilitation Program. The diagnostic assessment includes electromyography and manometry analyzed in 4 phases: (1) initial baseline phase; (2) rapid contraction phase; (3) tonic contraction and endurance phase; and (4) late baseline phase. This evaluation is performed at the onset of every session. PMR management consists of 6 possible therapeutic modalities, employed depending on the diagnostic evaluation: (1) down-training; (2) accessory muscle isolation; (3) discrimination training; (4) muscle strengthening; (5) endurance training; and (6) electrical stimulation. Eight to ten sessions are performed at one-week intervals with integration of home exercises and lifestyle modifications. Conclusions. The PMR protocol offers a standardized approach to diagnose and manage pelvic floor dysfunction syndromes with potential advantages over traditional biofeedback, involving additional interventions and a continuous pelvic floor assessment with management modifications over the clinical course.

Hydroxyapatite incorporated into collagen gels for mesenchymal stem cell culture

Collagen gels could be used as carriers in tissue engineering to improve cell retention and distribution in the defect. In other respect hydroxyapatite could be added to gels to improve mechanical properties and regulate gel contraction. The aim of this work was to analyze the feasibility to incorporate hydroxyapatite into collagen gels and culture mesenchymal stem cells inside it. Human bone marrow mesenchymal stem cells (hMSC-BM) were used in this study. Gels were prepared by mixing rat tail type I collagen, hydroxyapatite microparticles and MSCs. After polymerization gels were kept in culture while gel contraction and mechanical properties were studied. In parallel, cell viability and morphology were analyzed. Gels became free-floating gels contracted from day 3, only in the presence of cells. A linear rapid contraction phase was observed until day 7, then a very slow contraction phase took place. The incorporation of hydroxyapatite improved gel stability and mechanical properties. Cells were randomly distributed on the gel and a few dead cells were observed all over the experiment. This study shows the feasibility and biocompatibility of hydroxyapatite supplemented collagen gels for the culture of mesenchymal stem cells that could be used as scaffolds for cell delivery in osteoarticular regenerative medicine.

Flow structure and transport characteristics of feeding and exchange currents generated by upside-down Cassiopea jellyfish

Quantifying the flows generated by the pulsations of jellyfish bells is crucial for understanding the mechanics and efficiency of their swimming and feeding. Recent experimental and theoretical work has focused on the dynamics of vortices in the wakes of swimming jellyfish with relatively simple oral arms and tentacles. The significance of bell pulsations for generating feeding currents through elaborate oral arms and the consequences for particle capture are not as well understood. To isolate the generation of feeding currents from swimming, the pulsing kinematics and fluid flow around the benthic jellyfish Cassiopea spp. were investigated using a combination of videography, digital particle image velocimetry and direct numerical simulation. During the rapid contraction phase of the bell, fluid is pulled into a starting vortex ring that translates through the oral arms with peak velocities that can be of the order of 10 cm s(-1). Strong shear flows are also generated across the top of the oral arms throughout the entire pulse cycle. A coherent train of vortex rings is not observed, unlike in the case of swimming oblate medusae such as Aurelia aurita. The phase-averaged flow generated by bell pulsations is similar to a vertical jet, with induced flow velocities averaged over the cycle of the order of 1-10 mm s(-1). This introduces a strong near-horizontal entrainment of the fluid along the substrate and towards the oral arms. Continual flow along the substrate towards the jellyfish is reproduced by numerical simulations that model the oral arms as a porous Brinkman layer of finite thickness. This two-dimensional numerical model does not, however, capture the far-field flow above the medusa, suggesting that either the three-dimensionality or the complex structure of the oral arms helps to direct flow towards the central axis and up and away from the animal.

Limited Transplantation of Antigen-Expressing Hematopoietic Stem Cells Induces Long-Lasting Cytotoxic T Cell Responses

Harnessing the ability of cytotoxic T lymphocytes (CTLs) to recognize and eradicate tumor or pathogen-infected cells is a critical goal of modern immune-based therapies. Although multiple immunization strategies efficiently induce high levels of antigen-specific CTLs, the initial increase is typically followed by a rapid contraction phase resulting in a sharp decline in the frequency of functional CTLs. We describe a novel approach to immunotherapy based on a transplantation of low numbers of antigen-expressing hematopoietic stem cells (HSCs) following nonmyeloablative or partially myeloablative conditioning. Continuous antigen presentation by a limited number of differentiated transgenic hematopoietic cells results in an induction and prolonged maintenance of fully functional effector T cell responses in a mouse model. Recipient animals display high levels of antigen-specific CTLs four months following transplantation in contrast to dendritic cell-immunized animals in which the response typically declines at 4–6 weeks post-immunization. Majority of HSC-induced antigen-specific CD8+ T cells display central memory phenotype, efficiently kill target cells in vivo, and protect recipients against tumor growth in a preventive setting. Furthermore, we confirm previously published observation that high level engraftment of antigen-expressing HSCs following myeloablative conditioning results in tolerance and an absence of specific cytotoxic activity in vivo. In conclusion, the data presented here supports potential application of immunization by limited transplantation of antigen-expressing HSCs for the prevention and treatment of cancer and therapeutic immunization of chronic infectious diseases such as HIV-1/AIDS.

532 PRENATAL DEVELOPMENTAL HEALING WITH INTRA-ARTICULAR GROWTH HORMONE (IAGH) INJECTIONS TREATS A VARIETY OF OSTEOARTHRITIC AND RHEUMATOID JOINTS WITH A SUCCESS RATE OF 75% TO 95% AND NO COMPLICATIONS

plates. Gels without Hap were used as controls. The gels were kept in culture for 3 weeks in αMEM medium containing 10%FCS. Gel contraction and cell viability, morphology and metabolism were analyzed. Results: Gels, attached to surface plates after fabrication, became freefloating gels contracted from day 3, only in the presence of cells. A linear rapid contraction phase was observed until day 7, then a very slow contraction phase took place. The incorporation of Hap improved gel stability as compared to collagen gels without Hap. Total DNA increased two times from day 3 to day 21, reflecting cell proliferation inside gels. Moreover, while cell metabolism increased all over the culture period, an important increase was observed after 3 weeks. Cells cultured in gels showed elongated spindle-shape morphology and a few dead cells were observed all over the experiment. Microscopy images showed that cells were randomly distributed on the gel. After proliferation cells were connected, mainly at the gel surface, which could stabilize collagen contraction. Conclusions: This study shows the feasibility and biocompatibility of hydroxyapatite supplemented collagen gels for the culture of mesenchymal stem cells. They could be used as carriers for cell delivery in osteoarticular regenerative medicine.

Parvalbumin deficiency in fast‐twitch muscles leads to increased ‘slow‐twitch type’ mitochondria, but does not affect the expression of fiber specific proteins

Parvalbumin (PV), a small cytosolic protein belonging to the family of EF-hand calcium-binding proteins, is highly expressed in mammalian fast-twitch muscle fibers. By acting as a 'slow-onset' Ca2+ buffer, PV does not affect the rapid contraction phase, but significantly contributes to increase the rate of relaxation, as demonstrated in PV-/- mice. Unexpectedly, PV-/- fast-twitch muscles were considerably more resistant to fatigue than the wild-type fast-twitch muscles. This effect was attributed mainly to the increased fractional volume of mitochondria in PV-/- fast-twitch muscle, extensor digitorum longus, similar to levels observed in the slow-twitch muscle, soleus. Quantitative analysis of selected mitochondrial proteins, mitochondrial DNA-encoded cytochrome oxidase c subunit I and nuclear DNA-encoded cytochrome oxidase c subunit Vb and F1-ATPase subunit beta revealed the PV-/- tibialis anterior mitochondria composition to be almost identical to that in wild-type soleus, but not in wild-type fast-twitch muscles. Northern and western blot analyses of the same proteins in different muscle types and in liver are indicative of a complex regulation, probably also at the post-transcriptional level. Besides the function in energy metabolism, mitochondria in both fast- and slow-twitch muscles act as temporary Ca2+ stores and are thus involved in the shaping of Ca2+ transients in these cells. Previously observed altered spatio-temporal aspects of Ca2+ transients in PV-/- muscles are sufficient to up-regulate mitochondria biogenesis through the probable involvement of both calcineurin- and Ca2+/calmodulin-dependent kinase II-dependent pathways. We propose that 'slow-twitch type' mitochondria in PV-/- fast muscles are aimed to functionally replace the slow-onset buffer PV based on similar kinetic properties of Ca2+ removal.

Relative magnitude and asynchrony of directional components of contraction in sedated dog left ventricle

The purpose of this study was to quantitate the temporal relationships and the extent and speed of shortening in segments of myocardium responsive to contraction in circumferential, longitudinal, and oblique fiber groups. Measurements were made in five sedated dogs (morphine, diazepam) with and without alterations in preload and afterload (nitroprusside, phenylephrine). The measurement interval was the phase of rapid contraction, determined by differentiation of the segment length vs. time. In the control state, percentage segment shortening was greater in circumferential than in longitudinal [15.2 +/- 0.24 (SE) vs. 10.5 +/- 0.80%; P = 0.0020] and in the subepicardial oblique than in the subendocardial oblique fiber directions (16.6 +/- 0.65 vs. 9.7 +/- 0.36%; P = 0.0010). Shortening was proportional to both maximum speed and duration of shortening (r = 0.735 +/- 0.015 and 0.757 +/- 0.017, respectively). Duration of shortening was significantly longer in circumferential than in longitudinal (mean difference 39.3 +/- 6.6 ms; P = 0.0039) and in subepicardial oblique than in subendocardial oblique directions (mean difference 27.7 +/- 5.5 ms; P = 0.0072). Velocities of up to 3.0 segment lengths/s were attained in response to nitroprusside. These data reveal the local anisotropy and asynchrony of contraction in the myocardium; however, they also support the concept of the myocardium as a functional continuum. The dominance of circumferential over longitudinal and subepicardial over subendocardial oblique contractile components indicates their relative contributions to the constriction of the midmyocardial shell.

Tissue Selectivity and Calcium Dependence of Contractile Responses to Endothelin

The tissue selectivity and calcium dependence of endothelin, a potent vasoconstrictor peptide obtained from endothelial cells, were investigated in a number of rat and guinea pig tissues. Endothelin produced a significant dose-dependent contraction of rat tissues with the following rank order of efficacy: aorta greater than portal vein greater than trachea much greater than vas deferens = lung parenchyma much greater than bladder. In the rat atrium and ventricle, endothelin possessed positive inotropic effects at low doses and arrhythmogenic effects at high doses. Endothelin also produced a significant contraction of guinea pig lung parenchyma and induced relaxations of guinea pig taenia coli precontracted with potassium chloride. The aortic ring contractile response to endothelin was characterized by both rapid and slow phases of contraction. The rapid phase of contraction was abolished by removal of extracellular calcium or preincubation of tissues with the dihydropyridine calcium channel antagonist, nifedipine. Removal of the endothelium from aortic rings resulted in an increase of approximately 10-fold in the contractile potency of endothelin. In comparison, the potency of the dihydropyridine calcium channel activator, (-)-S-BAY K 8644, was increased approximately 15-fold by the same treatment. These findings suggest that endothelin, while displaying some selectivity as a vascular smooth muscle spasmogen, is a general smooth and cardiac muscle spasmogen which utilizes both intra- and extracellular sources of calcium to support its contractions.

Quantitative Evaluation of the Factors Affecting the Process of Fibroblast-Mediated Collagen Gel Contraction by Separating the Process into Three Phases

Kinetics of collagen gel contraction by fibroblasts cultured in vitro was examined in detail for quantitative analysis. The process of collagen gel contraction was not expressed by a simple function of time. It appeared to consist of three distinct phases; a lag phase before the initiation of contraction, a rapid contraction phase and a slow contraction phase. Factors affecting the gel contraction can be classified into four groups. The first group includes increase in cell number, in culture temperature or in serum concentration, which strengthened the contraction in all the three phases, suggesting that they affected cellular activity particularly in interacting with collagen. The second group repressed the later two phases of contraction but not the first lag phase, typically increase in collagen concentration and a low dose of nocodazole or colcemid. Increasing population doubling levels of fibroblasts belongs to the third group which caused a reduced lag time but no change in the later two phases. Cytochalasin D at a low dose (0.03–0.1 µg/ml) is another example of the third group which shortened the lag time. The last group did not change the contraction curves. Donor age of fibroblasts isolated from the skin is an example of this group. The rate of rapid contraction in the second phase was always found to be closely correlated with the degree of contraction at the end of the third phase, in a whole set of the factors above mentioned. The results suggest that the extent of the later two phases might be a reflection of the same cellular activity, particularly cytokinetical one. The lag time is directly related to the time for cells to become elongate in shape as observed by using the video-microscopy, suggesting that the lag phase is also governed by cytokinetical activity. The two cytokinetical activities are closely related, but may be distinct, since the factors affecting the collagen gel contraction can be differentiated into four groups.

Effects of an extract of Ginkgo biloba and diverse substances on the phasic and tonic components of the contraction of an isolated rabbit aorta

1. 1. The effects of phentolamine, propranolol, D 600, theophylline, papaverine and an extract of Ginkgo biloba were studied with respect to the two phases of the contractile response induced by norepinephrine in an isolated rabbit aorta. 2. 2. Phentolamine (3 × 10 −6 M) inhibits the rapid phase of the contraction of the rabbit aorta brought on by norepinephrine (NE) 10 −5 M more strongly than the tonic phase. Propranolol (10 −6 M) potentiates this rapid phase. 3. 3. D 600 inhibits the slow phase with an EC 50 = to 3.8 × 10 −8 M. 4. 4. Papaverine and theophylline increase the relaxation that follows the rapid phase of contraction. The slow phase is inhibited only by papaverine. 5. 5. The extract of Ginkgo biloba (Gb) at a concentration of 3 mg/ml has the same type of effect as papaverine 3 × 10 −5 M.

Actions of D 600 on isolated rabbit aorta

1. 1. The actions of D 600 (10 −9−10 −4 M) on various phases of NE- or Ca 2+-induced contractions of rabbit isolated aorta were determined in Ca 2+-free medium. 2. 2. D 600 inhibited the slow phase of contraction with EC 50 ≊ 3 × 10 −8 M , but affected the rapid phase of contraction only at 10 −4 M. 3. 3. D 600 increased the relaxation that followed the rapid phase of NE-induced contraction in Ca 2+-free medium with EC 50 ≊ 1.5 × 10 −5 M . 4. 4. The method described might be useful for further studies on Ca 2+-antagonists.

Inhibition of α-receptor-induced Ca 2+ release and Ca 2+ influx by Mn 2+ and La 3+

The influence of Mn 2+ and La 3+ on α-receptor-stimulated Ca 2+ movements was examined in arterial smooth muscle of the rabbit aorta. Both cations cause an inhibition of phenylephrine (PE) contractile response which exhibits a different pattern at low and high cation concentrations. At 0.1-1.0 nM inhibition by Mn 2+ and La 3+ was predominately due to a reduction in Ca 2+ influx reflected as inhibition of the slow phase of contraction and reduction in PE-stimulated 45Ca uptake. PE log dose-response curves were shifted to the right in a non-parallel manner by 1 mN Mn 2+ such that responses to lower PE concentrations were more inhibited. However, in the presence of 10 mM Mn 2+ PE responses are equally inhibited at all PE levels. At 10 mM both Mn 2+ and La 3+ also inhibited PE-stimulated Ca 2+ release resulting in a reduction in both the rapid phase of contraction and in the magnitude of PE stimulation of 45Ca efflux. The effects of Mn 2+ (1 or 10 mM) on contraction and 45Ca efflux were rapidly reversible, while the effect of La 3+ was not. Inhibition of Ca 2+ release by 10 mM Mn 2+ was not caused by displacement of releasable Ca 2+, but appeared to reflect their occupation of a superficially located receptor modulating site. The inhibition of Ca 2+ influx by lower concentrations of Mn 2+ may illustrate the functional consequence of configurational changes in the α 2-form of the receptor which have been recently described at lower concentrations of divalent cations.

Agonist induced release of intracellular Ca2+ in the rabbit aorta

The effects of hormonal agonists (norepinephrine, angiotensin, and histamine) on 45Ca efflux from the rabbit aorta were studied using a Ca-EGTA buffered efflux medium. Each caused a transient stimulation of efflux rate which probably reflected the release of an intracellular 45Ca store. The size of the stimulation of efflux correlated with the size of the initial rapid phase of contraction. The norepinephrine-sensitive intracellular Ca fraction was estimated to be greater than 21 mumoles/Kg wet tissue weight. This fraction is separate from intracellular Ca which is accumulated during relaxation. Evidence is presented for the lack of cyclic nucleotide involvement in the release of Ca2+, and possible alternative modes of coupling are discussed.

Premotion silent period in rapid voluntary movement.

The silent period in the agonist prior to rapid voluntary movement (premotion silent period) was observed in the lower limb muscles. The subject was asked to respond to the flashing light by performing a vertical jump as quickly as possible. The most consistent results were obtained in the extensors, particularly the quadriceps. The frequency of the silence of six subjects was 53% in the rectus femoris, 61% in the vastus medialis, and 72% in the vastus lateralis. A neural switching mechanism, directing excitation from preparation phase of jumping into rapid contraction phase, seemed to be attributable to the occurrence of the premotion silent period.

Aspects of stellar evolution reappraised

Abstract The theory of stellar evolution is reviewed in a simple fashion, with emphasis on topics for which some rethinking may be required. It is suggested that evolution of stars off the main sequence in the H-R diagram into the area occupied by the giants is caused, not by fuel exhaustion, but simply by instability following the ignition of thermonuclear energy release in the interior. The physics of a degenerate gas is discussed. It is pointed out that the equation of state remains the same as for a normal gas, but the pressure is exerted only by that fraction of the particles which occupy states within kT of the Fermi level. Degeneracy of the electron gas causes a large reduction of opacity, leading to an increased rate of loss of energy. The higher luminosity is fist fed by gravitational potential energy released during a phase of rapid contraction. However, when the proton Fermi level reaches a certain threshold, thermonuclear processes take over. During intial contraction angular momentum is trans...

Mode of action of sodium nitroprusside on vascular smooth muscle

1. Sodium nitroprusside is a potent relaxant of smooth muscles with a predominantly tonic response, e.g. rat aorta contracted by noradrenaline, angiotensin II, Phe2-Lys8-vasopressin, BaC1(2), or KC1, and guinea-pig tracheal smooth muscle contracted by carbachol. 2. Smooth muscle preparations from the splanchnic region and with varying degrees of phasic contractility are less sensitive and develop tachyphylaxis (portal vein, duodenum of the rat) or are unresponsive to sodium nitroprusside (vas deferens, uterus of the rat). 3. Cardiac auricles of the guinea pig are not affected by sodium nitroprusside in either frequency or amplitude or spontaneous contractions. 4. Sdium nitroprusside causes a parallel shift of the dose-response curve of rat aorta to noradrenaline to the right and reduces the maximum response. 5. The drug has no blocking or stimulant effect on alpha- or beta-adrenoceptors, respectively. 6. Sodium nitroprusside inhibits the contractile response of calcium-depleted depolarized rat aorta to extra-cellular calcium. Like verapamil, it inhibits the increment in 45calcium uptake of rabbit aorta elicited by K+. Sodium nitroprusside significantly reduced 45calcium binding by microsomes prepared from rabbit aorta. 7. Rabbit aorta was incubated with lanthanum chloride to prevent calcium influx; sodium nitroprusside reduced the maintained rapid contraction phase in response to noradrenaline which is believed to be based on the intracellular activation of calcium. 8. In rat aorta, cellular cAMP and ATP levels were not found to be affected by the drug. 9. Rabbit aorta, "skinned" by glycerination is unresponsive to sdoium nitroprusside. 10. It is concluded that sodium nitropruside acts on exictation-contraction coupling predominantly in tonic smooth muscle by interfering with both the influx and the intracellular activation of calcium.