Global Deletion of Pannexin 3 Resulting in Accelerated Development of Aging-Induced Osteoarthritis in Mice.

OBJECTIVE: Osteoarthritis (OA) results in pathologic changes in the joint tissue. The mechanisms driving disease progression remain largely unclear, and thus disease-modifying treatments are lacking. Pannexin 3 (Panx3) was identified as a potential mediator of cartilage degeneration in OA, and our previous study in mice indicated that deletion of the Panx3 gene delayed surgically induced cartilage degeneration. This study was undertaken to examine the role of Panx3 in other OA subtypes, particularly primary OA during aging, in a mouse model of aging-induced OA. METHODS: Wild-type (WT) and Panx3-/- C57BL/6J (Black-6) mice, ages 18-24 months, were analyzed by micro-computed tomography to investigate bone mineral density and body composition. Joints were harvested from the mice, and histopathologic analysis of the joint tissue for OA development was conducted with a specific focus on changes in articular cartilage, subchondral bone, and synovial tissue. RESULTS: Global loss of Panx3 in aging mice was not associated with increased mortality or changes in body composition. Mice lacking Panx3 had shorter appendicular skeletons than WT mice, but overall the body compositions appeared quite similar. Panx3 deletion dramatically accelerated cartilage degeneration and subchondral bone thickening with aging in both 18-month-old and 24-month-old mice, while promoting synovitis in 18-month-old mice. CONCLUSION: These observations in a mouse model of OA suggest that Panx3 has a protective role against the development of primary aging-associated OA. It appears that Panx3 has opposing context-specific roles in joint health following traumatic injury versus that associated with aging. These data strongly suggest that there are differences in the molecular pathways driving different subtypes of OA, and therefore a detailed understanding of these pathways could directly improve strategies for OA diagnosis, therapy, and research.

Mapping the twist-angle disorder and Landau levels in magic-angle graphene.

The recently discovered flat electronic bands and strongly correlated and superconducting phases in magic-angle twisted bilayer graphene (MATBG)1,2 crucially depend on the interlayer twist angle, θ. Although control of the global θ with a precision of about 0.1 degrees has been demonstrated1-7, little information is available on the distribution of the local twist angles. Here we use a nanoscale on-tip scanning superconducting quantum interference device (SQUID-on-tip)8 to obtain tomographic images of the Landau levels in the quantum Hall state9 and to map the local θ variations in hexagonal boron nitride (hBN)-encapsulated MATBG devices with relative precision better than 0.002 degrees and a spatial resolution of a few moiré periods. We find a correlation between the degree of θ disorder and the quality of the MATBG transport characteristics and show that even state-of-the-art devices-which exhibit correlated states, Landau fans and superconductivity-display considerable local variation in θ of up to 0.1 degrees, exhibiting substantial gradients and networks of jumps, and may contain areas with no local MATBG behaviour. We observe that the correlated states in MATBG are particularly fragile with respect to the twist-angle disorder. We also show that the gradients of θ generate large gate-tunable in-plane electric fields, unscreened even in the metallic regions, which profoundly alter the quantum Hall state by forming edge channels in the bulk of the sample and may affect the phase diagram of the correlated and superconducting states. We thus establish the importance of θ disorder as an unconventional type of disorder enabling the use of twist-angle gradients for bandstructure engineering, for realization of correlated phenomena and for gate-tunable built-in planar electric fields for device applications.

Src-type tyrosine kinase p56lck is critical for thymic stromal lymphopoietin-induced allergic rhinitis.

BACKGROUND: Thymic stromal lymphopoietin (TSLP) is a regulator of mast cell-mediated allergic inflammatory reactions, but the manner in which TSLP contributes to allergic rhinitis (AR) remains unclear. OBJECTIVE: Here, we sought to determine that TSLP plays a crucial role in AR by interacting with Src-type tyrosine kinase p56lck and STAT6 and promoting mast cells degranulation. METHODS: The effects of TSLP on mast cell degranulation and AR were analysed in human mast cell line (HMC-1 cells), ovalbumin (OVA)-induced AR animal model, and human subjects. Small interfering RNA experiments were performed in HMC-1 cells and OVA-induced AR model. Immune responses were analysed by enzyme-linked immunosorbent assay, Western blotting, immunoprecipitation, and histological studies. RESULTS: Thymic stromal lymphopoietin levels and mast cell-derived p56lck activation were elevated in human subjects with AR, and in AR mice, exogenous TSLP accelerated TH2-allergic inflammatory reactions by up-regulating p56lck and STAT6. On the other hand, depletion of TSLP, p56lck, and STAT6 ameliorated clinical symptoms in AR mice. The selective inhibitor of p56lck, damnacanthal, inhibits AR reactions. CONCLUSION: Collectively, these observations suggest a role for TSLP/p56lck/STAT6 in AR and offer insight into potential therapeutic strategies.

Evaluation of flexural, diametral tensile, and shear bond strength of composite repairs.

OBJECTIVE: Repairing composite restorations may be a more conservative treatment than replacing the entire restoration. The objective of this in vitro study was to determine the best repair method by measuring flexural, diametral tensile, and shear bond strength of repaired composites in which the surfaces were treated with chemical primers (Add & Bond or Silane Bond Enhancer), a bonding agent (Optibond Solo Plus [OBSP]), or mechanical retention with a bonding agent. METHODS: Filtek Supreme Ultra shade B1B was placed in special molds to fabricate specimens that served to test the flexural, diametral tensile, or shear strength of the inherent resin substrate. The same molds were modified to make specimens for testing repair strength of the resin. Repairs were made immediately or after aging in deionized water at 37°C for seven days. All repair sites were finished with coarse Sof-Lex discs to simulate finishing new restorations or partially removing aged restorations. Repair surfaces were treated with one of the following: 1) phosphoric-acid etching and OBSP; 2) Add & Bond; 3) phosphoric-acid etching, Silane Bond Enhancer, and OBSP; or 4) quarter round bur, phosphoric-acid etching, and OBSP. Specimens were placed back in the original molds to fabricate specimens for diametral tensile or flexural testing or in an Ultradent jig to make specimens for shear bond testing. Composite resin in shade B5B was polymerized against the treated surfaces to make repairs. Two negative control groups for the three testing methods consisted of specimens in which repairs were made immediately or after aging without any surface treatments. Controls and experimental repairs were aged (water 37°C, 24 hours) before flexural, diametral tensile, or shear testing in an Instron Universal testing machine at a crosshead speed of 0.5 mm/min. RESULTS: Experimental flexural repair strengths ranged from 26.4% to 88.6% of the inherent substrate strength. Diametral tensile repair strengths ranged from 40% to 80% of the inherent substrate strength, and shear bond strength repairs ranged from 56% to 102%. Geometric means were statistically analyzed with two-way analysis of variance on their log-transformed values. Significant differences were determined using Tukey honestly significant difference (p<0.05). CONCLUSIONS: Depending on the mechanical property being tested, surface treatments produced different results. OBSP produced more consistent results than chemical primers.

Fractal images induce fractal pupil dilations and constrictions.

Fractals are self-similar structures or patterns that repeat at increasingly fine magnifications. Research has revealed fractal patterns in many natural and physiological processes. This article investigates pupillary size over time to determine if their oscillations demonstrate a fractal pattern. We predict that pupil size over time will fluctuate in a fractal manner and this may be due to either the fractal neuronal structure or fractal properties of the image viewed. We present evidence that low complexity fractal patterns underlie pupillary oscillations as subjects view spatial fractal patterns. We also present evidence implicating the autonomic nervous system's importance in these patterns. Using the variational method of the box-counting procedure we demonstrate that low complexity fractal patterns are found in changes within pupil size over time in millimeters (mm) and our data suggest that these pupillary oscillation patterns do not depend on the fractal properties of the image viewed.

Hofstadter's butterfly and the fractal quantum Hall effect in moiré superlattices.

Electrons moving through a spatially periodic lattice potential develop a quantized energy spectrum consisting of discrete Bloch bands. In two dimensions, electrons moving through a magnetic field also develop a quantized energy spectrum, consisting of highly degenerate Landau energy levels. When subject to both a magnetic field and a periodic electrostatic potential, two-dimensional systems of electrons exhibit a self-similar recursive energy spectrum. Known as Hofstadter's butterfly, this complex spectrum results from an interplay between the characteristic lengths associated with the two quantizing fields, and is one of the first quantum fractals discovered in physics. In the decades since its prediction, experimental attempts to study this effect have been limited by difficulties in reconciling the two length scales. Typical atomic lattices (with periodicities of less than one nanometre) require unfeasibly large magnetic fields to reach the commensurability condition, and in artificially engineered structures (with periodicities greater than about 100 nanometres) the corresponding fields are too small to overcome disorder completely. Here we demonstrate that moiré superlattices arising in bilayer graphene coupled to hexagonal boron nitride provide a periodic modulation with ideal length scales of the order of ten nanometres, enabling unprecedented experimental access to the fractal spectrum. We confirm that quantum Hall features associated with the fractal gaps are described by two integer topological quantum numbers, and report evidence of their recursive structure. Observation of a Hofstadter spectrum in bilayer graphene means that it is possible to investigate emergent behaviour within a fractal energy landscape in a system with tunable internal degrees of freedom.

Massive Dirac fermions and Hofstadter butterfly in a van der Waals heterostructure.

van der Waals heterostructures constitute a new class of artificial materials formed by stacking atomically thin planar crystals. We demonstrated band structure engineering in a van der Waals heterostructure composed of a monolayer graphene flake coupled to a rotationally aligned hexagonal boron nitride substrate. The spatially varying interlayer atomic registry results in both a local breaking of the carbon sublattice symmetry and a long-range moiré superlattice potential in the graphene. In our samples, this interplay between short- and long-wavelength effects resulted in a band structure described by isolated superlattice minibands and an unexpectedly large band gap at charge neutrality. This picture is confirmed by our observation of fractional quantum Hall states at ± 5/3 filling and features associated with the Hofstadter butterfly at ultrahigh magnetic fields.

Evaluating dentin surface treatments for resin-modified glass ionomer restorative materials.

This in vitro study evaluated the effect of six surface treatments on the shear bond strength of three resin-modified glass ionomers (RMGIs) to dentin. Occlusal surfaces of caries-free third molars were reduced to expose only dentin. Surface treatments were smear layer intact (negative control), Cavity Conditioner, EDTA, Ketac Primer, Self Conditioner, and etching with 35% phosphoric acid followed by the application of Optibond Solo Plus. Filtek Z250 composite resin bonded with Optibond Solo Plus served as a positive control. Conditioning agents were used according to the manufacturers' instructions. After surface treatments, Fuji II LC, Riva LC, Ketac Nano, and Filtek Z250 were placed in copper-band matrices 5 mm in diameter and 2 mm in height and were light-cured for 20 seconds. Specimens were stored in 100% humidity for 24 hours, after which they were placed in deionized water for 24 hours at 37°C. They were then tested under shear forces in an Instron Universal Testing Machine at a crosshead speed of 0.5 mm/min. A two-way analysis of variance and Tukey honestly significant difference statistical analyses (p<0.05) indicated significant interaction between RMGIs and conditioning agents. Acid etching followed by Optibond Solo Plus provided highest bond strengths for all three RMGIs, which were not statistically different from the positive control.

Activation of hypoxia-inducible factor-1 regulates human histidine decarboxylase expression.

Histidine decarboxylase (HDC) catalyzes the formation of histamine from histidine. Histamine has various effects in physiological and pathological reactions, such as inflammation, cell growth, and neuro-transmission. We investigated the role of hypoxia-inducible factor (HIF)-1 on hypoxia-induced HDC expression in human mast cell line, HMC-1 cells and mouse bone marrow-derived mast cells (BMMCs). Hypoxia significantly increased histamine production. HDC expression and activity were induced by hypoxia. Additionally, when cells were transfected with a native form of HIF-1alpha, hypoxia could induce higher HDC expression than in the nontransfected cell. HIF-1 binding activity for HDC 5' flanking region (HFR) was similar to that for the hypoxia-responsive element. Using HDC promoter deletion analysis, we also demonstrated that HFR was regulated by HIF-1 activation. In addition, depletion of HIF-1alpha prevents hypoxic induction of HDC in BMMCs. In conclusion, these results demonstrate that hypoxia induces HDC expression by transcriptional mechanisms dependent upon HIF-1.

Alginic acid has anti-anaphylactic effects and inhibits inflammatory cytokine expression via suppression of nuclear factor-kappaB activation.

BACKGROUND: Alginic acid is comprised of complex polymerized polysaccharides, and can be chemically extracted from seaweed. Alginic acid has an inhibitory effect on histamine release, but its molecular mechanisms are not well understood. OBJECTIVE: To investigate the effect of alginic acid on the mast cell-mediated anaphylactic and inflammatory reaction using in vivo and in vitro models and elucidate its molecular mechanisms. MATERIALS AND METHOD: The effect of alginic acid on an allergy model was analysed by anaphylaxis, a histidine decarboxylase (HDC) assay, and a histamine assay. Cytokine production was analysed by means of ELISA. Cytokine expression was analysed by means of RT-PCR, and Western blotting. Transcription factor activity was analysed by a luciferase assay and a transcription factor-enzyme linked immunoassay. RESULTS: Alginic acid dose dependently inhibited compound 48/80-induced systemic anaphylaxis with doses of 0.25-1 g/kg 1 h (P<0.01, n=6) and significantly inhibited passive cutaneous anaphylaxis by 54.8%. Alginic acid (0.01-1 microg/mL) inhibited histamine release from serum and peritoneal mast cells (P<0.01). All these effects were stronger than those of disodium cromoglycate (DSCG), the reference drug tested. Alginic acid also inhibited HDC expression and activity on the phorbol myristate acetate (PMA)+A23187-stimulated human mast cell line, HMC-1 cells. Moreover, alginic acid significantly inhibited the production of PMA+A23187-induced inflammatory cytokines, IL-1beta and TNF-alpha, but not that of IL-6 or IL-8. In activated HMC-1 cells, the expression level of nuclear factor (NF)-kappaB/Rel A protein increased in the nucleus, whereas the level of NF-kappaB/Rel A in the nucleus was decreased by alginic acid treatment. In addition, alginic acid (0.01 microg/mL) decreased the PMA+A23187-induced luciferase activity and DNA-binding activity. CONCLUSION: The present results indicate that alginic acid has potent anti-anaphylactic and anti-inflammatory properties.

Fetal oxygen content is restored after maternal hemorrhage and fluid replacement with polymerized bovine hemoglobin, but not with hetastarch, in pregnant sheep.

UNLABELLED: We investigated the ability of hemoglobin-based oxygen carrying solutions (HBOCs) to alleviate fetal hypoxemia from maternal hemorrhage. Fifteen pregnant ewes (132-day gestational age) were hemorrhaged 20 mL/kg over 1 h; they were randomized to receive 20 mL/kg IV of HBOC, hetastarch (HTS), or autologous blood (BLD) (n = 5 each) over 30 min and were monitored for 2 h. Hemorrhage significantly (P < or = 0.05) decreased maternal mean blood pressure (from 98 to 48 mm Hg, median), arterial oxygen content (from 12.2 to 11.1 mL/dL), and fetal arterial oxygen content (from 8.1 to 3.9 mL/dL). Fluid replacement restored maternal blood pressure in all groups, although maternal oxygen content immediately returned to baseline only after BLD or HBOC. Maternal oxygen saturation decreased after HBOC (from 98% to 88%). Fetal oxygen content rapidly returned to baseline with either BLD (7.1 mL/dL) or HBOC (8.0 mL/dL) but was never restored with HTS (4.7 mL/dL), and, 60 min after fluid replacement, it was higher with HBOC (8.3 mL/dL) than with HTS (4.7 mL/dL). Fetal plasma-free hemoglobin did not change after HBOC. In conclusion, maternal fluid replacement with HBOC or BLD effectively restored fetal oxygenation, primarily by restoring maternal oxygen content, whereas HTS did not. IMPLICATIONS: Hemoglobin solutions eliminate many limitations of blood transfusions. Our results show that fluid replacement with either blood or a hemoglobin solution, compared with hetastarch, restored fetal oxygenation in pregnant ewes after hemorrhage. If applicable to women, these results suggest a potential for the use of hemoglobin solutions in obstetrics.

In vitro comparison of the fracture resistance and failure mode of fiber, ceramic, and conventional post systems at various stages of restoration.

PURPOSE: This in vitro study evaluated 6 post systems over 4 simulated clinical stages of tooth restoration to (1) determine quantitatively the fracture resistance strength at each stage when a static loading force is applied to cause failure; (2) determine the failure mode for each post system at each simulated clinical stage; and (3) determine the feasibility of removing failed post systems. MATERIALS AND METHODS: Ten post systems made with various materials and designs were tested at the following 4 stages of simulated clinical treatment: stage #1: posts only, loaded using a 3-point loading model to failure, to determine transverse strengths and failure modes for each post system; stage #2: posts alone, bonded into teeth; stage #3: posts bonded into teeth with core build up; stage #4: post and core build up and full veneer restoration. For stages #2 through #4, the coronal portion of 60 mandibular premolars was amputated at the cemento-enamel junction, the canals were treated endodontically, and the specimens were mounted in acrylic blocks. A testing force was applied to the posts at 90 degrees to the long axis of the tooth, 4 mm from the cemento-enamel junction. The O'Brien test for constant variance was performed over the treatment groups. For nonconstant variance, the Welsh analysis of variance was used to test for equalities of treatment means. The Tukey Kramer procedure determined which treatment procedures differed. RESULTS: The failure thresholds for each post system were significantly different at each stage of testing, but the order of test results by post type remained generally consistent from one stage to the next. ParaPosts (Coltene, Whaledent Int, New York, NY) and core build up resulted in significantly higher failure thresholds through all 4 stages of testing. This post system also consistently displayed a high number of nonfavorable tooth fractures. FibreKor post and cores (Jeneric Pentron Inc, Wallingford, CT) resulted in significantly lower failure threshold values in stages #2 through #4. This post system displayed no tooth fractures in stages #2 and #3 and a similar number of nonfavorable tooth fractures in stage #4 when compared with the other systems. C-Post (Bisco Dental Products, Schaumburg, IL), CosmoPost (Ivoclar Vivadent North America Inc, Amherst, NY), and AesthetiPost (Bisco Dental Products) grouped in descending order through stages #2 to #4. These systems displayed intermediate fracture resistance strengths, as well as a moderate number of nonfavorable tooth fractures. CosmoPost exhibited a significant number of brittle post fractures with fragments left in the root canal at all stages. The fracture resistance of the cast metal post varied from stage to stage. No teeth fractured at stage #2. At stage #3, 9 of 10 teeth fractured nonfavorably, and all teeth fractured nonfavorably in stage #4. CONCLUSIONS: The fiber posts evaluated provided an advantage over a conventional post that showed a higher number of irretrievable post and unrestorable root fractures. At the stage of final restoration insertion, there was no difference in force to failure for all but the FibreKor material, which continued to be weaker than all other tested materials. The fiber posts were readily retrievable after failure, whereas the remaining post systems tested were nonretrievable. J Prosthodont 2001;10:26-36.

Periparturient and neonatal anesthesia.

Small animal patients may need to be anesthetized in the periparturient period for emergency, nonobstetric reasons, elective ovariohysterectomy, or cesarean section. In each case, the physiologic changes in the dam must be accounted for in designing an anesthetic protocol, but the requirements of the fetuses will be different. Subsequent to birth, the neonatal animal may need to be anesthetized, and the unique physiology and pharmacology at this age is described.

Neonatal critical care.

The first few minutes after a neonate's birth may determine the quality of its entire life. Immediate care includes prevention of hypothermia, clearing of nasal and oral passages, stimulation of ventilation and oxygenation, and, in a few cases, advanced life support. Any additional stress during the first weeks of life can also result in neonatal morbidity and mortality. Care of the diseased newborn must focus not only on treatment of the underlying disease but on aggressive supportive care. A safe, warm, clean, proper environment and adequate nutrition are essential.

Perioperative risk factors for puppies delivered by cesarean section in the United States and Canada.

The purpose of this study was to evaluate perioperative risk factors affecting neonatal survival after cesarean section. Data from 807 cesarean-derived litters (3,908 puppies) was submitted by 109 practices in the United States and Canada. Survival rates immediately, two hours, and seven days after delivery were 92% (n=3,127), 87% (n=2,951), and 80% (n=2,641), respectively, for puppies delivered by cesarean section (n=3,410) and were 86% (n=409), 83% (n=366), and 75% (n=283), respectively, for puppies born naturally (n=498). Maternal mortality rate was 1% (n=9). Of 776 surgeries, 453 (58%) were done on an emergency basis. The most common breed of dog was bulldog (n=138; 17%). The most common methods of inducing and maintaining anesthesia were administration of isoflurane for induction and maintenance (n=266; 34%) and administration of propofol for induction followed by administration of isoflurane for maintenance (n=237; 30%). A model of cesarean-derived puppies surviving to birth, between birth and two hours, and between two hours and seven days was designed to relate litter survival to perioperative factors. The following factors increased the likelihood of all puppies being alive: the surgery was not an emergency; the dam was not brachycephalic; there were four puppies or less in the litter; there were no naturally delivered or deformed puppies; all puppies breathed spontaneously at birth; at least one puppy vocalized spontaneously at birth; and neither methoxyflurane nor xylazine was used in the anesthetic protocol.

Unilateral thalamic stroke does not decrease ipsilateral sleep spindles.

STUDY OBJECTIVES: To measure the sleep spindle characteristics in patients with unilateral thalamic stroke. DESIGN: A prospective study of patients with thalamic stroke and age-matched healthy controls. SETTING: Department of Neurology of a University Hospital. PARTICIPANTS: Thirteen patients (mean age: 67 years, SD: 13,44) with an isolated, unilateral acute thalamic stroke and 18 healthy age-matched volunteers. INTERVENTIONS: A polysomnogram recording from 14 scalp EEG electrodes performed during 2 consecutive nights, the second or third week after the stroke. Only the sleep of the second night was analyzed. MEASUREMENTS AND RESULTS: Sleep spindles were counted during two separate 10-minute epochs of stage II. Spindles appearing synchronously in both sides with similar amplitude were called "bilateral." Spindles with twice the amplitude in one side than the other were "right" or "left-side predominant". There were 8 patients with posterolateral, 3 with global and 2 with anterior lesions. Eight were right and 5 left-sided. The number of spindles was similar in patients (39.8 +/- 23.4 in 20 minutes) than controls (26.07 +/- 29.07; p=0.173). Spindles with a centroparietal (34%) and centroparieto-occipital localization (22%) were the most frequent. In controls approximately 66% of the spindles had a bilateral and symmetric distribution over the scalp, 23% of the spindles were predominantly left-sided and 5% were predominantly right-sided. In patients, bilateral spindles decreased (p<0.0001) but asymmetric spindles did not change. CONCLUSION: Unilateral acute thalamic stroke does not decrease sleep spindles ipsilaterally; rather, it seems to produce a bilateral diminution in their number.