Smooth Muscle Cells Of Blood Vessels Research Articles

A case of giant cell osteosarcoma in a Scottish fold cat

This study describes a case of giant cell osteosarcoma (GCO) localised on the right scapulae in a six-year-old neutered male Scottish Fold cat. GCO is a rare tumour in domestic animals. This tumour is also known as giant cell-rich osteosarcoma. Multinucleated giant cells and osteoid structures are observed in GCO. The mass was 4.6 x 4.4 x 4.3 cm in size and very firm. Microscopically, fusiform spindle-shaped cells and multinucleated giant cells were observed. There were 4-8 mitotic figures in three random high-power fields (400x). Osteoid structures and necrosis were detected in several areas. In these areas, atypia was observed in the cells. Immunohistochemically, vimentin expression was observed in neoplastic cells and multinucleated giant cells. No immunoreactivity against actin and cytokeratin was observed in neoplastic cells. Additionally, vimentin expression was detected in the periphery of blood vessels and actin expression in blood vessel smooth muscle cells. As a result of histopathological and immunohistochemical findings, the mass was determined to be GCO.

Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis.

Synovitis of the knee synovium is proven to be a precursor of knee osteoarthritis (OA), leading to a radiologically advanced stage of the disease. This study was conducted to elucidate the expression pattern of different inflammatory factors—NF-kB, iNOS, and MMP-9 in a subpopulation of synovial cells. Thirty synovial membrane intra-operative biopsies of patients (ten controls, ten with early OA, and ten with advanced OA, according to the Kellgren–Lawrence radiological score) were immunohistochemically stained for NF-kB, iNOS, and MMP9, and for different cell markers for macrophages, fibroblasts, leukocytes, lymphocytes, blood vessel endothelial cells, and blood vessel smooth muscle cells. The total number of CD68+/NF-kB+ cells/mm2 in the intima of early OA patients (median = 2359) was significantly higher compared to the total number of vimentin+/Nf-kB+ cells/mm2 (median = 1321) and LCA+/NF-kB+ cells/mm2 (median = 64) (p < 0.001 and p < 0.0001, respectively). The total number of LCA+/NF-kB+ cells/mm2 in the subintima of advanced OA patients (median = 2123) was significantly higher compared to the total number of vimentin+/NF-kB+ cells/mm2 (median = 14) and CD68+/NF-kB+ cells/mm2 (median = 29) (p < 0.0001). The total number of CD68+/iNOS+ cells/mm2 in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/iNOS+ cells/mm2 and LCA+/iNOS+ cells/mm2 (p < 0.0001 and p < 0.001, respectively). The total number of CD68+/MMP-9+ cells/mm2 in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/MMP-9+ cells/mm2 and CD5+/MMP-9+ cells/mm2 (p < 0.0001). Macrophages may have a leading role in OA progression through the NF-kB production of inflammatory factors (iNOS and MMP-9) in the intima, except in advanced OA, where leukocytes could have a dominant role through NF-kB production in subintima. The blocking of macrophageal and leukocyte NF-kB expression is a possible therapeutic target as a disease modifying drug.

A Mini Review on Hemostasis and Coagulation

Hemostasis, the cycle by which draining is captured, includes a mind boggling arrangement of physiological and biochemical occasions that end in the development of a steady attachment that seals the vein. The cycle includes connection between the vein divider and platelets, blood coagulation, and fibrinolysis. The endothelium assumes a significant function in obsessive cycles, for example, atherosclerosis, apoplexy, and scattered intravascular coagulation, flawed hemostasis, irritation, invulnerable issues, vascular neoplasia, and metastasis. Following injury, there is expanded entanglement of plasma lipoprotein parts followed by multiplication of blood vessel smooth muscle cells and additionally testimony of free cholesterol and cholesterol esters, anew union of connective tissue, and intracellular and extracellular affidavit of lipids in factor sums. Then again, coagulation continues by an inborn or intravascular pathway and by an outward or tissue juice pathway, the two of which convert prothrombin to thrombin. The vast majority of the properties and elements of Hageman factor apply to plasma thromboplastin predecessor, which additionally assumes a significant part in the underlying initiation phases of coagulation.

A Note on Adenocarcinoma

Hemostasis, the cycle by which draining is captured, includes a mind boggling arrangement of physiological and biochemical occasions that end in the development of a steady attachment that seals the vein. The cycle includes connection between the vein divider and platelets, blood coagulation, and fibrinolysis. The endothelium assumes a significant function in obsessive cycles, for example, atherosclerosis, apoplexy, and scattered intravascular coagulation, flawed hemostasis, irritation, invulnerable issues, vascular neoplasia, and metastasis. Following injury, there is expanded entanglement of plasma lipoprotein parts followed by multiplication of blood vessel smooth muscle cells and additionally testimony of free cholesterol and cholesterol esters, anew union of connective tissue, and intracellular and extracellular affidavit of lipids in factor sums. Then again, coagulation continues by an inborn or intravascular pathway and by an outward or tissue juice pathway, the two of which convert prothrombin to thrombin [1]. The vast majority of the properties and elements of Hageman factor apply to plasma thromboplastin predecessor, which additionally assumes a significant part in the underlying initiation phases of coagulation.

Cellular Distribution of Canonical and Putative Cannabinoid Receptors in Canine Cervical Dorsal Root Ganglia.

Growing evidence indicates cannabinoid receptors as potential therapeutic targets for chronic pain. Consequently, there is an increasing interest in developing cannabinoid receptor agonists for treating human and veterinary pain. To better understand the actions of a drug, it is of paramount importance to know the cellular distribution of its specific receptor(s). The distribution of canonical and putative cannabinoid receptors in the peripheral and central nervous system of dogs is still in its infancy. In order to help fill this anatomical gap, the present ex vivo study has been designed to identify the cellular sites of cannabinoid and cannabinoid-related receptors in canine spinal ganglia. In particular, the cellular distribution of the cannabinoid receptors type 1 and 2 (CB1 and CB2) and putative cannabinoid receptors G protein-coupled receptor 55 (GPR55), nuclear peroxisome proliferator-activated receptor alpha (PPARα), and transient receptor potential vanilloid type 1 (TRPV1) have been immunohistochemically investigated in the C6–C8 cervical ganglia of dogs. About 50% of the neuronal population displayed weak to moderate CB1 receptor and TRPV1 immunoreactivity, while all of them were CB2-positive and nearly 40% also expressed GPR55 immunolabeling. Schwann cells, blood vessel smooth muscle cells, and pericyte-like cells all expressed CB2 receptor immunoreactivity, endothelial cell being also PPARα-positive. All the satellite glial cells (SGCs) displayed bright GPR55 receptor immunoreactivity. In half of the study dogs, SGCs were also PPARα-positive, and limited to older dogs displayed TRPV1 immunoreactivity. The present study may represent a morphological substrate to consider in order to develop therapeutic strategies against chronic pain.

Effect of 27nt-miRNA on the differentiation of mesenchymal stem cells into vascular smooth muscle cells

To investigate the effect of 27nt-miRNA on the differentiation of mesenchymal stem cells into vascular smooth muscle cells. The highly expression plasmids of 27nt-miRNA and anti-27nt-miRNA, and negative control plasmids were constructed, packaged with lentivirus and transfected into human umbilical cord mesenchymal stem cells (hUCMSCs). Collagen IV was added to induce hUCMSCs differentiation into blood vessel smooth muscle cells (VSMCs). The cell viability was measured by MTT assay. The expression of SMA, SM22α at mRNA and protein levels was determined by RT-PCR, immunocytochemical staining and Western blotting. Compared with the negative control group, the viability of the 27nt-miRNA overexpression group was decreased by 20.48% (P<0.05), and the expression of SMA mRNA and SM22α mRNA and protein was significantly increased (P<0.05); the viability of Anti-27nt-miRNA group was increased 18.07% (P<0.05), and the expression of SMA mRNA and SM22α mRNA and protein was decreased (P<0.05). In summary, 27nt-miRNA promotes mesenchymal stem cells differentiation into vascular smooth muscle cells and inhibits cells viability.

The Effect of Voltage‐Sensitive Chloride Channel 6 on Development of Salt‐Sensitive Hypertension

Hypertension is currently the number one risk factor for global disease burden. Advancing our knowledge on the underlying mechanisms of blood pressure (BP) control is essential to reduce the incidence of hypertension. Recent human genome‐wide association studies have linked a number of rare coding mutations and intronic single nucleotide polymorphisms in the gene, CLCN6, to lower average systolic and diastolic blood pressure as well as reduced hypertension risk. The voltage‐gated chloride channel 6 (CLC6) is encoded by CLCN6, which is one of 9 voltage‐gated chloride channel (CLC) family members. The functions of CLC6 at molecular, cellular, and physiological levels are largely unknown.We have previously introduced a 5 amino acid deletion into Clcn6 in the Dahl Salt Sensitive (SS) rat background (SS‐Clcn6em2Mcwim (hereafter called CLC6 KO)). These rats demonstrated lower mean arterial pressure (MAP) compared to WT SS rats (Flister et al., Genome Res, 2013) when placed on a high salt (HS, 4% NaCl) diet for 10 days, suggesting these animals might serve as a model system to study the role of CLCN6 in modulating blood pressure. To further test the role of CLC6 on blood pressure and renal function, we placed CLC6 KO or WT SS rats on a HS diet for 3 weeks and recorded their blood pressure using telemetry. Electrolytes and kidney injury were also analyzed. CLC6 KO rats had a significantly lower BP maintaining a MAP about 15 mmHg lower than the WT SS rats. There were no significant differences in plasma Na+, K+, Cl−, or Ca2+ levels between WT SS and CLC6 KO rats on NS diets; however, the plasma K+ levels of the CLC6 KO rats on the HS diet were significantly elevated compared to controls. Immunohistochemical analysis revealed that CLC6 is strongly expressed in blood vessel smooth muscle cells. Therefore, we studied vasodilatory response of isolated mesenteric resistance arteries in response to increasing concentrations of acetylcholine. We found that arteries from the CLC6 KO animals had impaired dilation to 10 μM acetylcholine, and relaxed to 25 ± 2.9 % of their maximal diameter relative to controls (60 ± 13.9 %). Lastly, we measured spontaneous intracellular Ca2+ activity in intact ex vivo mesenteric artery smooth muscle at single cell resolution using event detection analysis of two‐photon microscopy videos. Cells were constricted with phenylephrine then induced to relax with acetylcholine, and Ca2+ cycling was significantly reduced in the CLC6 KO (19.0 ± 1.7 sec vs 93.0 ± 11.3 sec) during relaxation, consistent with the vessel dilation studies.These results suggest that CLC6 KO is protective against development of salt sensitive hypertension by altering the intracellular Ca2+ activity in arterial smooth muscle cells, but additional studies are needed to more fully understand this effect.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Overview of Triptans in the Treatment of Acute Migraine

The advent of triptans for effective relief of migraine represented a therapeutic breakthrough. Triptans are serotonin (5-hydroxytryptamine, or 5-HT) agonists with high affinity for 5-HT1B and 5-HT1D receptors. There are, at present, seven commonly used triptans: almotriptan, eletriptan, frovatriptan, naratriptan, rizatriptan, sumatriptan, and zolmitriptan. Some controversy still surrounds the mode of action of this class. When first studied, it was thought that triptans provided relief from migraine through cranial vasoconstriction, probably via action at postsynaptic 5-HT1B receptors on the smooth-muscle cells of blood vessels. More recently, however, triptans have also been demonstrated to block release of vasoactive peptides from the perivascular trigeminal neurons owing to their action at presynaptic 5-HT1D receptors on the nerve terminal. Triptans may also facilitate descending pain inhibitory systems. However, it is not certain whether or not the activation of vascular 5-HT1B receptors is essential for relieving migraine. Many drug characteristics need to be taken into account when selecting the best triptan for an individual patient. Clinical characteristics of the migraine attack and the patient’s lifestyle and medical history are also important. Despite their biochemical similarity, triptans have distinct pharmacokinetic and pharmacodynamic profiles. Frovatriptan and naratriptan, for example, have a longer half-life and therefore a delayed onset of action and prolonged duration compared with the other triptans, which are fast acting, with a rapid dose-dependent efficacy and higher risk of adverse events and migraine recurrence. Migraine recurrence is affected by the pharmacological and pharmacokinetic properties of the triptan but is unrelated to initial clinical efficacy. Triptans with a longer half-life and largest 5-HT1B receptor affinity have the lowest rates of headache recurrence.

Modulatory role of nitric oxide in cardiac performance

Nitric oxide is produced by almost all cardiac cells, endothelial cells, cardiomyocytes and nerve fibers. It is synthesized by an enzyme, a nitric oxide synthase, which occurs in endothelial, neural and inducible form. The distribution of nitric oxide synthase in the heart is characterized by a pronounced non-uniformity. Nitric oxide exerts its effects in physiological and pathophysiological conditions. The physiological effects of low concentrations of nitric oxide, which is released in the normal conditions under the influence of constituent enzymes, occur via cyclic guanosine monophosphate. The synthesized nitric oxide exhibits its effect in the cells where it is produced, in an autocrine manner, or by diffusing into the neighboring cells, in a paracrine manner. Nitric oxide acts by regulating the coronary vessel tonus, affecting the contractility of cardiomyocytes, generating an inotropic effect in a dose-dependent manner and controlling the cellular respiration. Other effects of nitric oxide in the cardiovascular system include the hyperpolarization of the smooth muscle cells in blood vessels, the inhibition of the monocyte adhesion, the inhibition of platelet migration, adhesion and aggregation and the proliferation of smooth muscle cells and fibroblasts. The anti-atherosclerotic effects of nitric oxide are based on these effects. Nitric oxide is a weak free radical in gaseous state, and the cytotoxic and/or the cytoprotective effects of the higher concentrations of nitric oxide are related to the chemical structure of nitric oxide as a free radical. The excessive production of nitric oxide by the activation of inducible nitric oxide synthase can lead to major irregularities in the function of cardiomyocytes and cardiac insufficiency. Understanding the nitric oxide molecular mechanisms of signaling pathways in the heart can provide a new strategic approach to prevention and treatment of cardiovascular diseases.

Fatty Acid Binding Protein 1 Is Related with Development of Aspirin-Exacerbated Respiratory Disease

BackgroundAspirin-exacerbated respiratory disease (AERD) refers to the development of bronchoconstriction in asthmatics following the ingestion of aspirin. Although alterations in eicosanoid metabolites play a role in AERD, other immune or inflammatory mechanisms may be involved. We aimed to identify proteins that were differentially expressed in nasal polyps between patients with AERD and aspirin-tolerant asthma (ATA).Methodology/Principal FindingsTwo-dimensional electrophoresis was adopted for differential display proteomics. Proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS). Western blotting and immunohistochemical staining were performed to compare the amount of fatty acid-binding protein 1 (FABP1) in the nasal polyps of patients with AERD and ATA. Fifteen proteins were significantly up- (seven spots) or down-regulated in the nasal polyps of patients with AERD (n = 5) compared to those with ATA (n = 8). LC-MS revealed an increase in seven proteins expression and a decrease in eight proteins expression in patients with AERD compared to those with ATA (P = 0.003–0.045). FABP1-expression based on immunoblotting and immunohistochemical analysis was significantly higher in the nasal polyps of patients with AERD compared to that in patients with ATA. FABP1 was observed in epithelial, eosinophils, macrophages, and the smooth-muscle cells of blood vessels in the polyps.Conclusions/SignificanceOur results indicate that alterations in 15 proteins, including FABP1, may be related to the development of AERD.

Synthesis, characterizations and biocompatibility of alternating block polyurethanes based on P3/4HB and PPG‐PEG‐PPG

Block copolymers with exactly controlled structures, that is, alternating block polyurethanes based on poly(3-hydroxybutyrate-co-4-hydroxybutrate) (P3/4HB-diol) and poly (propylene glycol)-poly(ethylene glycol)-poly(propylene glycol) (PPG-PEG-PPG) were synthesized by solution polymerization via specifically selective coupling reaction between terminal hydroxyl P3/4HB segment and isocyanate group end-capped PPG-PEG-PPG segment, using 1,6-hexamethylene diisocyanate (HDI) as end-capped agent. The chemical structure, molecular weight and distribution were systematically characterized by nuclear magnetic resonance spectrum (¹H NMR), Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC). The thermal property was investigated by differential scanning calorimetry (DSC) and thermogravimetric analysis. The hydrophilicity was studied by static contact angle of H₂O and CH₂I₂. DSC revealed that the PU3/4HB-alt-PPG-PEG-PPG exhibited a distinct change from amorphous to 30% crystallinity degree, T(g) from -25 to -50 °C, T(m) from 110 to 145 °C. The polyurethanes were more hydrophilic (water contact angle centers around 80 °) than the raw PHA materials. The platelet adhesion assay showed that the obtained polyurethanes had a lower platelet adhesion than the raw materials and the amount of platelet adhesion could be controlled by varying the segmental length of P3/4HB-diol. This could be explained by the inclusion of PPG-PEG-PPG between the P3/4HB segments, improving the hemocompatibility of P3/4HB. The cell culture assay revealed that the obtained polyurethanes were cell inert and unfavorable for the attachment of mouse fibroblast cell line L929 and rabbit blood vessel smooth muscle cells (RaSMCs). This suggests that these polyurethanes would be promising candidates as hemocompatibility and tissue-inert materials.

Diffuse intraneural leiomyoma in a case of sensorimotor neuropathy

Leiomyomas are benign smooth muscle tumors, usually characterized by well-deWned Wbrous pseudocapsule and intersecting fascicles of spindle cells [3]. Few cases of leiomyomas in the peripheral nervous system (PNS) causing external nerve compression have been described [1, 7]. We report the case of a 71-year-old man who was referred for a 2 years history of burning pain and paresthesias associated with slowly progressive weakness of the lower limbs. He underwent total colectomy for diVuse polyposis 2 years before. Neurological examination and nerve conduction studies revealed symmetric sensorimotor axonal neuropathy of the lower limbs. Since all immunological, hematological and cerebrospinal Xuid examinations were negative, a sural nerve biopsy was performed. On semithin plastic sections, a mild degree of endoneurial Wbrosis, reduction of myelinated Wbers and signs of acute axonal degeneration were evident. Some fascicles had greater myelinated Wber loss than others, primarily in the center of the fascicles. Interestingly, paraYn-embedded, hematoxylin and eosinstained sections showed in epineurial and endoneurial perivascular spaces atypical large cells with abundant eosinophilic cytoplasm. Upon electron microscopy, these cells exhibited a basement membrane and prominent pinocytotic vesicles at the periphery (Fig. 1). The cytoplasm was Wlled with actin and myosin Wlaments, aggregates of glycogen particles and few organelles. Some cells showed oval and centrally located nuclei. Immunohistochemically, they were positive for smooth muscle actin, desmin and calponin (Fig. 2), but negative for CD31, cytokeratin and S-100 protein, demonstrating that they were smooth muscle cells (SMC). These cells did not show proliferative activity (negativity for Ki-67). Neither necrosis nor a deWnite capsule were present. The morphological and immunohistochemical data suggested a diagnosis of leiomyoma. This is the Wrst description of a diVuse SMC inWltrate of the PNS, both in the endoneurium and epineurium, whereas well circumscribed and encapsulated leiomyomas have occasionally been reported in the PNS. Other potential tumors such as leiomyosarcoma, schwannoma and hemangioma have to be considered in the diVerential diagnosis, but can be excluded based on immunoproWle [2]. Several hypotheses can be envisaged for the possible cellular origin of this unique tumor in the PNS, including pluripotential mesenchymal cells, perivascular connective tissue elements or SMC of blood vessels [6]. Epineurial proliferation of SMC in sural nerve biopsies indicating inXammatory or non-inXammatory angiopathy has been described before [5]. However, our patient’s sural nerve biopsy revealed SMC also in the endoneurium without signs of angiopathy, suggesting a diVerent origin of SMC. How this inWltrate caused damage of peripheral nerve Wbers remains unclear. Our patient’s clinical presentation suggests a diVuse damage of the PNS, while nerve biopsy Wndings associated with F. Cerri (&) Department of Neurology and INSPE, San RaVaele ScientiWc Institute, Via Olgettina, 6

The expression of cross-linked elastin by rabbit blood vessel smooth muscle cells cultured in polyhydroxyalkanoate scaffolds

In order to evaluate the possibility for making artificial blood vessels, a series of microbial copolyesters poly(3-hydroxybutyrate- co-4-hydroxybutyrate)s (P3HB4HB)s containing 0–40 mol% 4-hydroxybutyrate (4HB) were studied for growth and formation of elastin of rabbit blood vessel smooth muscle cells (RaSMCs) incubated on the solvent-casting polyester films. Porous scaffolds of all P3HB4HBs except P(HB-40 mol% 4HB) were used to compare their potentials as materials for tissue engineering blood vessel (TEBV). The polyesters were studied using static contact angles, differential scanning calorimetry (DSC), cell count kit-8 (CCK-8) and Fastin elastin assays. Simultaneously, SEM, H&E and DAPI staining were employed to study cell morphology and cell growth in the polyester scaffolds. Viability of rabbit blood vessel smooth muscle cells (RaSMCs) grown on P(HB-7 mol% 4HB) films was the strongest among all other tested polyester films during the whole growth process. H&E and DAPI staining clearly suggested good cells' growth and even confluent growth in the P3HB4HB scaffolds. Fastin elastin assay demonstrated that 4HB component in the P3HB4HB copolymer benefited the elastin formation and accumulation. P(HB-20 mol% 4HB) showed a 160% more elastin production compared with that on the well-studied poly(3-hydroxybutyrate- co-12 mol% 3-hydroxyhexanoate) (PHBHHx) incubated under the same conditions. Since the P3HB4HB has adjustable elasticity and strength, combined with its ability to induce elastin formation, it can be regarded as a useful material for developing into a material for artificial blood vessels.

Third‐generation dihydropyridine‐type calcium channel blocker labedipinedilol‐B displays α/β‐adrenoceptor blocking activities

AbstractThe pharmacological properties of labedipinedilol‐B {N‐[4‐[2‐hydroxy‐3‐(2‐methoxy‐1‐oxyethylaminobenzene) propoxy]‐benzyl]‐2,6‐dimethyl‐3,5‐dicarbomethoxy‐1,4‐dihydropyridine} were investigated in vivo and in vitro in comparison with labedipinedilol‐A. Intravenous labedipinedilol‐B (0.5, 1.0, and 3.0 mg kg–1), produced dose‐dependent hypotensive and bradycardia responses in pentobarbital‐anesthetized Wistar rats. Pretreatment with labedipinedilol‐B (1.0 mg kg–1, iv) also inhibited phenylephrine (10 μg kg–1)‐induced hypertensive and (–)isoproterenol (0.5 μg kg–1)‐induced tachycardia effects. In the isolated Wistar rat right and left atria and guinea pigs tracheal strips experiments, labedipinedilol‐B (10–7, 10–6, and 10–5 M) competitively antagonized the (–)isoproterenol‐induced positive chronotropic and inotropic effects and tracheal relaxation responses in a concentration‐dependent manner. The parallel shift to the right of the concentration–response curve of (–)isoproterenol suggested that labedipinedilol‐B was a β1/β2‐adrenoceptor competitive antagonist. Labedipinedilol‐B (10–7, 10–6, and 10–5 M) also prevented the rate‐increasing effects of increased extracellular Ca2+ (3.0–9.0 mM) in a concentration‐dependent manner. In the isolated rat aorta, labedipinedilol‐B (10–7, 10–6, and 10–5 M) competitively antagonized the CaCl2 and norepinephrine‐induced contractions with pKCa–1 and pA2 values of 8.02 ± 0.04 and 7.55 ± 0.05 in a concentration‐dependent manner. The parallel shift to the right of the concentration–response curves of norepinephrine suggested that labedipinedilol‐B was an α‐adrenoceptor competitive antagonist. Furthermore, labedipinedilol‐B, in an equal antagonist activity, inhibited norepinephrine‐induced phasic and tonic contraction. In the isolated rat aorta, labedipinedilol‐B also competitively antagonized CaCl2‐induced contractions and made the parallel shift to the right of the concentration–response curve of CaCl2. In cultured blood vessel smooth muscle cells (A7r5 cell lines), Bay K 8644‐induced intracellular calcium changes were decreased after application of labedipinedilol‐B, suggesting that the compound was a calcium channel blocker. The binding characteristics of labedipinedilol‐B were evaluated in [3H]CGP‐12177 binding to ventricle and lung and [3H]prazosin binding to brain membranes in rats. Labedipinedilol‐B also was evaluated in [3H]nitrendipine binding to brain membranes in rats. These results indicated that labedipinedilol‐B, similar to labedipinedilol‐A, has α‐adrenoceptor blocking, β‐adrenoceptor blocking, and calcium entry blocking activities in a single compound. We suggest that these two compounds represent a new generation of 1,4‐dihydropyridine‐type calcium channel blockers. Drug Dev. Res. 52:462–474, 2001. © 2001 Wiley‐Liss, Inc.

Dual expression and differential regulation of phosphodiesterase 3A and phosphodiesterase 3B in human vascular smooth muscle: implications for phosphodiesterase 3 inhibition in human cardiovascular tissues.

Cyclic nucleotide phosphodiesterases (PDEs) are a superfamily of enzymes whose physiological role is the attenuation of the signaling mediated by the ubiquitous second messengers cAMP and cGMP. Given the myriad of physiological processes regulated by cAMP and cGMP, PDEs have long been studied as potential therapeutic targets. Although phosphodiesterase 3 (PDE3) activity is abundant in human cardiovascular tissues, and acute PDE3 inhibition, with agents such as milrinone, was beneficial in heart failure patients, prolonged treatments were associated with time-dependent reductions in hemodynamic effects and increased mortality. The molecular basis of this time-dependent reduction in efficacy has not been elucidated. In this context, we used a combination of approaches to determine PDE3 expression in human cardiovascular tissues and to elucidate the effects of prolonged elevations of cellular cAMP, as would occur with PDE3 inhibition, on this activity. Although our data confirms the expression of PDE3A in human blood vessel smooth muscle cells (HASMCs), we identify a previously unrecognized role for PDE3B in cAMP hydrolysis in human cardiovascular tissues. Specifically, although both PDE3A and PDE3B were expressed in HASMCs, their subcellular expression pattern and regulated expression by cAMP were distinct, with only expression of PDE3B being subject to cAMP-regulated expression. Thus, a paradigm emerges that allows for dual expression, with distinctive regulation, of both PDE3A and PDE3B proteins in cardiovascular tissues that may have profound significance for the rational design of molecules regulating this PDE activity.

Expression of intermediate filament proteins in fetal and adult human lung tissues

Abstract The expression patterns of intermediate filament proteins in fetal and normal or nonpathological adult human lung tissues are described using (chain-specific) monoclonal antibodies. In early stages of development (9–10 weeks and 25 weeks of gestation) only so-called simple cytokeratins such as cytokeratins 7 (minor amounts), 8, 18 and 19 are detected in bronchial epithelial cells. At later stages of development, the cytokeratin expression patterns become more complex. The number of bronchial cells positive for cytokeratin 7 increases, but basal cells in the bronchial epithelium remain negative. These latter cells show, however, expression of cytokeratin 14 in the third trimester of gestation. Developing alveolar epithelial cells express cytokeratins 7, 8, 18 and 19. In adult human bronchial epithelium cytokeratins 4 (varying amounts), 7, 8, 13 (minor amounts), 14, 18 and 19 can be detected, with the main expression of cytokeratins 7, 8, and 18 in columnar cells and the main expression of cytokeratin 14 in basal cells. Vimentin is detected in all mesenchymal tissues. In addition, fetal lung expresses vimentin in bronchial epithelium, however, to a lesser extent with increasing age, resulting in the expression of vimentin in only few scattered bronchial cells at birth. Also in adult bronchial epithelium the expression of vimentin is noticed in part of the basal and columnar epithelial cells. Desmin filaments, present in smooth muscle cells of the lung, appear to alter their protein structure with age. In early stages of development smooth muscle cells surrounding blood vessels are partly reactive with some cytokeratin antibodies and with a polyclonal desmin antibody. At week 9–10 and week 25 of gestation a monoclonal antibody to desmin, however, is not reactive with blood vessel smooth muscle cells but is only reactive with smooth muscle cells surrounding bronchi. With increasing age the reactivity of cytokeratin antibodies with smooth muscle cells in blood vessels decreases, while the reactivity with the monoclonal desmin antibody increases. Our results show that duroing differentation profound changes in the intermediate filament expression patterns occur in the different cell types of the developing lung.

Otologic Histopathology of Fabry's Disease

Fabry's disease is a rare progressive X-linked recessive disorder of glycosphingolipid metabolism. The accumulation of glycosphingolipids occurs in virtually all areas of the body, including the endothelial, perithelial, and smooth-muscle cells of blood vessels, the ganglion cells of the autonomic nervous system, and the glomeruli and tubules of the kidney. Although otologic symptoms have been described in these patients, to our knowledge there have been no temporal bone histopathologic reports. We describe the clinical histories, audiometric results, and temporal bone findings of two patients with this rare disorder. Both patients demonstrated a bilateral sloping sensorineural hearing loss audiometrically. Middle ear findings of seropurulent effusions and hyperplastic mucosa were seen in all four temporal bones. Strial and spiral ligament atrophy in all turns, and hair cell loss mainly in the basal turns, were also common findings. The number of spiral ganglion cells was reduced in all temporal bones; however, evidence of glycosphingolipid accumulation was not observed in the spiral ganglia.