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1.
Phys Rev Lett ; 130(18): 183602, 2023 May 05.
Article in English | MEDLINE | ID: mdl-37204878

ABSTRACT

The Heisenberg limit to laser coherence C-the number of photons in the maximally populated mode of the laser beam-is the fourth power of the number of excitations inside the laser. We generalize the previous proof of this upper bound scaling by dropping the requirement that the beam photon statistics be Poissonian (i.e., Mandel's Q=0). We then show that the relation between C and sub-Poissonianity (Q<0) is win-win, not a tradeoff. For both regular (non-Markovian) pumping with semiunitary gain (which allows Q→-1), and random (Markovian) pumping with optimized gain, C is maximized when Q is minimized.

2.
BMC Neurol ; 21(1): 355, 2021 Sep 14.
Article in English | MEDLINE | ID: mdl-34521381

ABSTRACT

BACKGROUND: Continuous spike and wave of sleep with encephalopathy (CSWS) is a rare and severe developmental electroclinical epileptic encephalopathy characterized by seizures, abundant sleep activated interictal epileptiform discharges, and cognitive regression or deceleration of expected cognitive growth. The cause of the cognitive symptoms is unknown, and efforts to link epileptiform activity to cognitive function have been unrevealing. Converging lines of evidence implicate thalamocortical circuits in these disorders. Sleep spindles are generated and propagated by the same thalamocortical circuits that can generate spikes and, in healthy sleep, support memory consolidation. As such, sleep spindle deficits may provide a physiologically relevant mechanistic biomarker for cognitive dysfunction in epileptic encephalopathies. CASE PRESENTATION: We describe the longitudinal course of a child with CSWS with initial cognitive regression followed by dramatic cognitive improvement after treatment. Using validated automated detection algorithms, we analyzed electroencephalograms for epileptiform discharges and sleep spindles alongside contemporaneous neuropsychological evaluations over the course of the patient's disease. We found that sleep spindles increased dramatically with high-dose diazepam treatment, corresponding with marked improvements in cognitive performance. We also found that the sleep spindle rate was anticorrelated to spike rate, consistent with a competitively shared underlying thalamocortical circuitry. CONCLUSIONS: Epileptic encephalopathies are challenging electroclinical syndromes characterized by combined seizures and a deceleration or regression in cognitive skills over childhood. This report identifies thalamocortical circuit dysfunction in a case of epileptic encephalopathy and motivates future investigations of sleep spindles as a biomarker of cognitive function and a potential therapeutic target in this challenging disease.


Subject(s)
Brain Diseases , Diazepam , Child , Cognition , Electroencephalography , Humans , Sleep
3.
J Mol Med (Berl) ; 99(2): 289-301, 2021 02.
Article in English | MEDLINE | ID: mdl-33392634

ABSTRACT

Primary brain tumors remain among the deadliest of all cancers. Glioma grade IV (glioblastoma), the most common and malignant type of brain cancer, is associated with a 5-year survival rate of < 5%. Melatonin has been widely reported as an anticancer molecule, and we have recently demonstrated that the ability of gliomas to synthesize and accumulate this indolamine in the surrounding microenvironment negatively correlates with tumor malignancy. However, our understanding of the specific effects mediated through the activation of melatonin membrane receptors remains limited. Thus, here we investigated the specific roles of MT1 and MT2 in gliomas and medulloblastomas. Using the MT2 antagonist DH97, we showed that MT1 activation has a negative impact on the proliferation of human glioma and medulloblastoma cell lines, while MT2 activation has an opposite effect. Accordingly, gliomas have a decreased mRNA expression of MT1 (also known as MTNR1A) and an increased mRNA expression of MT2 (also known as MTNR1B) compared to the normal brain cortex. The MT1/MT2 expression ratio negatively correlates with the expression of cell cycle-related genes and is a positive prognostic factor in gliomas. Notably, we showed that functional selective drugs that simultaneously activate MT1 and inhibit MT2 exert robust anti-tumor effects in vitro and in vivo, downregulating the expression of cell cycle and energy metabolism genes in glioma stem-like cells. Overall, we provided the first evidence regarding the differential roles of MT1 and MT2 in brain tumor progression, highlighting their relevance as druggable targets. KEY MESSAGES: • MT1 impairs while MT2 promotes the proliferation of glioma and medulloblastoma cell lines. • Gliomas have a decreased expression of MT1 and an increased expression of MT2 compared to normal brain cortex. • Tumors with a high MT1/MT2 expression ratio have significantly better survival rates. • Functional selective drugs that simultaneously activate MT1 and inhibit MT2 downregulate the expression of cell cycle and energy metabolism genes in glioma stem-like cells and exert robust anti-tumor effects in vivo.


Subject(s)
Brain Neoplasms , Glioma , Receptor, Melatonin, MT1 , Receptor, Melatonin, MT2 , Animals , Brain/metabolism , Brain Neoplasms/genetics , Brain Neoplasms/metabolism , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation , Disease Progression , Female , Glioma/genetics , Glioma/metabolism , Glioma/mortality , Glioma/pathology , Humans , Kaplan-Meier Estimate , Male , Mice, Inbred BALB C , Mice, Nude , Receptor, Melatonin, MT1/genetics , Receptor, Melatonin, MT1/metabolism , Receptor, Melatonin, MT2/genetics , Receptor, Melatonin, MT2/metabolism
4.
Obes Sci Pract ; 5(1): 46-58, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30847225

ABSTRACT

OBJECTIVE: To determine the efficacy of a 4-month school-based health, nutrition and exercise intervention on body fatness and examine possible effects of demographic and anthropometric covariates. METHODS: Height, weight, waist circumference and body composition were measured in a diverse population of 644 NYC middle school students (mean ± SD age 12.7 ± 0.9 years; 46% male; 38% Hispanic, 17% East Asian, 15% South Asian, 13.5% African American, 8.5% Caucasian, 8% other) during the fall and spring semesters. Year 1 participants (n = 322) were controls. Experimental participants (year 2, n = 469) received a 12-session classroom-based health and nutrition educational programme with an optional exercise intervention. RESULTS: Groups were demographically and anthropometrically similar. The intervention resulted in significant reductions in indices of adiposity (ΔBMI z-scores [-0.035 ± 0.014; p = 0.01], Δ% body fat [-0.5 ± 0.2; p < 0.0001] and Δwaist circumference [-0.73 ± 0.30 cm; p < 0.0001]). Intervention effects were greater (p = 0.01) in men (ΔBMI z-score = -0.052 ± 0.015) versus women (0.022 ± 0.018), participants who were obese (ΔBMI z-score -0.083 ± 0.022 kg m-2) versus lean (-0.0097 ± 0.020 kg m-2) and South Asians (Δ% body fat -1.03 ± 0.35) versus total (-0.49 ± 0.20%) participants (p = 0.005). CONCLUSION: A 4-month school-based health intervention was effective in decreasing measures of adiposity in middle school students, particularly in men, participants who were obese and South Asians.

5.
Nanoscale ; 9(15): 4907-4917, 2017 Apr 13.
Article in English | MEDLINE | ID: mdl-28358158

ABSTRACT

The solid concentration of pulmonary mucus (wt%) is critical to respiratory health. In patients with respiratory disease, such as Cystic Fibrosis (CF) and Chronic Obstructive Pulmonary Disorder (COPD), mucus hydration is impaired, resulting in high wt%. Mucus with high wt% is a hallmark of pulmonary disease that leads to obstructed airways, inflammation, and infection. Methods to measure mucus hydration in situ and in real-time are needed for drug development and personalized therapy. We employed plasmonic gold nanorod (GNR) biosensors that intermittently collide with macromolecules comprising the mucus mesh as they self-diffuse, such that GNR translational diffusion (DT) is sensitive to wt%. GNRs are attractive candidates for bioprobes due to their anisotropic optical scattering that makes them easily distinguishable from native tissue using polarization-sensitive OCT. Using principles of heterodyne dynamic light scattering, we developed diffusion-sensitive optical coherence tomography (DS-OCT) to spatially-resolve changing DT in real-time. DS-OCT enables, for the first time, direct monitoring of changes in nanoparticle diffusion rates that are sensitive to nanoporosity with spatial and temporal resolutions of 4.7 µm and 0.2 s. DS-OCT therefore enables us to measure spatially-resolved changes in mucus wt% over time. In this study, we demonstrate the applicability of DS-OCT on well-differentiated primary human bronchial epithelial cells during a clinical mucus-hydrating therapy, hypertonic saline treatment (HST), to reveal, for the first time, mucus mixing, cellular secretions, and mucus hydration on the micrometer scale that translate to long-term therapeutic effects.


Subject(s)
Biosensing Techniques , Epithelial Cells/cytology , Gold , Mucus/chemistry , Nanotubes , Bronchi/cytology , Cells, Cultured , Diffusion , Humans , Lung Diseases/drug therapy , Tomography, Optical Coherence
6.
Gene Ther ; 21(3): 253-61, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24451115

ABSTRACT

Primary ciliary dyskinesia (PCD) is a genetically heterogenous autosomal recessive disease in which mutations disrupt ciliary function, leading to impaired mucociliary clearance and life-long lung disease. Mouse tracheal cells with a targeted deletion in the axonemal dynein intermediate chain 1 (Dnaic1) gene differentiate normally in culture but lack ciliary activity. Gene transfer to undifferentiated cultures of mouse Dnaic1(-/-) cells with a lentiviral vector pseudotyped with avian influenza hemagglutinin restored Dnaic1 expression and ciliary activity. Importantly, apical treatment of well-differentiated cultures of mouse Dnaic1(-/-) cells with lentiviral vector also restored ciliary activity, demonstrating successful gene transfer from the apical surface. Treatment of Dnaic1(flox/flox) mice expressing an estrogen-responsive Cre recombinase with different doses of tamoxifen indicated that restoration of ∼20% of ciliary activity may be sufficient to prevent the development of rhinosinusitis. However, although administration of a ß-galactosidase-expressing vector into control mice demonstrated efficient gene transfer to the nasal epithelium, treatment of Dnaic1(-/-) mice resulted in a low level of gene transfer, demonstrating that the severe rhinitis present in these animals impedes gene transfer. The results demonstrate that gene replacement therapy may be a viable treatment option for PCD, but further improvements in the efficiency of gene transfer are necessary.


Subject(s)
Axonemal Dyneins/metabolism , Ciliary Motility Disorders/therapy , Genetic Therapy , Lentivirus/genetics , Animals , Axonemal Dyneins/genetics , Cells, Cultured , Cilia/metabolism , Cilia/physiology , Estrogen Antagonists/pharmacology , Genetic Vectors/genetics , Integrases/genetics , Lentivirus/metabolism , Mice , Mice, Inbred C57BL , Respiratory Mucosa/drug effects , Respiratory Mucosa/metabolism , Rhinitis/therapy , Sinusitis/therapy , Tamoxifen/pharmacology
7.
Transplant Proc ; 44(10): 2910-2, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23194995

ABSTRACT

BACKGROUND: The determination of kidney function plays a pivotal role in living donors renal assessment because of the long-term hazards of life with one kidney. Guidelines recommend estimation of glomerular filtration rate (GFR) by the Modification of Renal Disease (MDRD) or Cockroft-Gault equations for people with normal or near-normal renal function. Cystatin C (CysC) has been introduced as an alternative endogenous marker of GFR. OBJECTIVE: The objective of the study was to evaluate residual renal function among living kidney donors by comparing serum CysC concentrations and estimated GFR according to the MDRD formula or the Cockroft-Gault equation. PATIENTS AND METHODS: Forty living kidney donors showed a mean age of 46.14 years. Their GFR was estimated according to the abbreviated MDRD (aMDRD) and Cockroft-Gault formula adjusted for body surface area. Twenty-two donors underwent diethylenetriaminepentaacetic acid (DTPA) renal studies. Serum CysC concentrations were measured during the last follow-up visit. GFR values according to Cockroft-Gault formula and MDRD formula were correlated with CysC concentrations using Pearson's linear correlation. RESULTS: Mean GFR according to the aMDRD formula and Cocroft-Gault formula decreased after nephrectomy. The Cockroft-Gault formula overestimated the DTPA GFR in our study. No significant differences were observed between DTPA GFR and GFR estimated using the aMDRD equation. The rate of GFR decrease was approximately 0.8 mL/min/1.73 m(2) per year. No significant correlation was observed between serum CysC concentration and GFR. Microalbuminuria was observed in one patient after nephrectomy. CONCLUSIONS: aMDRD equation to estimate GFR is more precise than Cockroft-Gault formula and cystatin C in living kidney donors after nephrectomy and should be preferred model in these patients.


Subject(s)
Cystatin C/blood , Glomerular Filtration Rate , Kidney Transplantation/methods , Kidney/physiopathology , Living Donors , Nephrectomy , Tissue and Organ Harvesting/methods , Adult , Aged , Biomarkers/blood , Female , Humans , Kidney/diagnostic imaging , Kidney/metabolism , Kidney Transplantation/adverse effects , Linear Models , Male , Middle Aged , Models, Biological , Nephrectomy/adverse effects , Predictive Value of Tests , Radionuclide Imaging , Radiopharmaceuticals , Technetium Tc 99m Pentetate , Tissue and Organ Harvesting/adverse effects , Treatment Outcome
8.
Am J Physiol Lung Cell Mol Physiol ; 302(2): L238-47, 2012 Jan 15.
Article in English | MEDLINE | ID: mdl-22003093

ABSTRACT

The relationships between airway epithelial Cl(-) secretion-Na(+) absorption balance, airway surface liquid (ASL) homeostasis, and lung disease were investigated in selected transgenic mice. 1) To determine if transgenic overexpression of wild-type (WT) human CFTR (hCFTR) accelerated Cl(-) secretion and regulated Na(+) absorption in murine airways, we utilized a Clara cell secretory protein (CCSP)-specific promoter to generate mice expressing airway-specific hCFTR. Ussing chamber studies revealed significantly (∼2.5-fold) elevated basal Cl(-) secretory currents in CCSP-hCFTR transgenic mouse airways. Endogenous murine airway Na(+) absorption was not regulated by hCFTR, and these mice exhibited no lung disease. 2) We tested whether hCFTR, transgenically expressed on a transgenic mouse background overexpressing the ß-subunit of the epithelial Na(+) channel (ß-ENaC), restored ion transport balance and ASL volume homeostasis and ameliorated lung disease. Both transgenes were active in CCSP-hCFTR/ß-ENaC transgenic mouse airways, which exhibited an elevated basal Cl(-) secretion and Na(+) hyperabsorption. However, the airway disease characteristic of ß-ENaC mice persisted. Confocal studies of ASL volume homeostasis in cultured tracheal cells revealed ASL autoregulation to a height of ∼6 µm in WT and CCSP-hCFTR cultures, whereas ASL was reduced to <4 µm in ß-ENaC and CCSP-hCFTR/ß-ENaC cultures. We conclude that 1) hCFTR overexpression increases basal Cl(-) secretion but does not regulate Na(+) transport in WT mice and 2) transgenic hCFTR produces increased Cl(-) secretion, but not regulation of Na(+) channels, in ß-ENaC mouse airways and does not ameliorate ß-ENaC mouse lung disease.


Subject(s)
Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Epithelial Sodium Channels/metabolism , Ion Transport/genetics , Lung Diseases/metabolism , Respiratory Mucosa/metabolism , Sodium Channels/metabolism , Animals , Cells, Cultured , Chlorides/metabolism , Epithelial Sodium Channels/genetics , Genotype , Lung/metabolism , Lung Diseases/genetics , Mice , Mice, Inbred C57BL , Mice, Transgenic , Promoter Regions, Genetic , Respiratory Mucosa/cytology , Respiratory Mucosa/pathology , Respiratory System , Sodium/metabolism , Sodium Channels/genetics , Trachea/metabolism , Uteroglobin/genetics
9.
Am J Physiol Cell Physiol ; 296(6): C1301-9, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19321738

ABSTRACT

The nasal epithelium of the cystic fibrosis (CF) mouse has been used extensively in CF research because it exhibits ion transport defects similar to those of human CF airways. This tissue is composed of approximately 50% olfactory (OE) and approximately 50% ciliated epithelium (CE), and on the basis of previous observations, we hypothesized that a significant fraction of the bioelectric signals from murine nasal tissue may arise from OE rather than CE, while CE is the target tissue for CF gene therapy. We compared the bioelectric properties of isolated OE from the nasal cavity and CE from the nasopharynx in Ussing chamber studies. Hyperabsorption of Na(+) [amiloride response; CF vs. wild type (WT)] was approximately 7.5-fold greater in the OE compared with the CE. The forskolin response in native tissues did not reliably distinguish genotypes, likely due to a cyclic nucleotide-gated cation conductance in OE and a calcium-mediated Cl(-) conductance in CE. By potential difference assay, hyperabsorption of Na(+) (CF vs. WT) and the difference in response to apical 0 Cl(-) buffer (CF vs. WT) were approximately 2-fold greater in the nasal cavity compared with the nasopharynx. Our studies demonstrate that in the CF mouse, both the hyperabsorption of Na(+) and the Cl(-) transport defect are of larger magnitude in the OE than in the CE. Thus, while the murine CF nasal epithelium is a valuable model for CF studies, the bioelectrics are likely dominated by the signals from the OE, and assays of the nasopharynx may be more specific for studying the ciliated epithelium.


Subject(s)
Chlorides/metabolism , Cystic Fibrosis/metabolism , Nasal Cavity/metabolism , Nasopharynx/metabolism , Olfactory Mucosa/metabolism , Respiratory Mucosa/metabolism , Sodium/metabolism , Age Factors , Amiloride/pharmacology , Animals , Cilia/metabolism , Colforsin/pharmacology , Disease Models, Animal , Humans , Ion Transport , Male , Membrane Potentials , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CFTR , Nasal Cavity/drug effects , Nasopharynx/drug effects , Olfactory Mucosa/drug effects , Respiratory Mucosa/drug effects , Tissue Culture Techniques
10.
Gene Ther ; 14(20): 1492-501, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17637798

ABSTRACT

Successful gene therapy will require that the therapeutic gene be expressed at a sufficient level in the correct cell type(s). To improve the specificity of gene transfer for cystic fibrosis (CF) and other airway diseases, we have begun to develop cell-type specific promoters to target the expression of transgenes to specific airway cell types. Using a FOXJ1 promoter construct previously shown to direct transgene expression specifically to ciliated cells, we have generated transgenic mice expressing human cystic fibrosis transmembrane conductance regulator (CFTR) in the murine tracheal and nasal epithelia. RNA analysis demonstrated levels of CFTR expression is greater than or equal to the level of endogenous mouse CFTR. Immunoprecipitation and western blotting demonstrated the production of human CFTR protein, and immunochemistry confirmed that CFTR was expressed in the apical region of ciliated cells. The transgenic animals were bred to CFTR null mice (Cftr(tm1Unc)) to determine if expression of CFTR from the FOXJ1 promoter is capable of correcting the airway defects in Cl(-) secretion and Na(+) absorption that accompany CF. Isolated trachea from neonatal CF mice expressing the FOXJ1/CFTR transgene demonstrated a correction of forskolin-stimulated Cl(-) secretion. However, expression of human CFTR in ciliated cells of the nasal epithelia failed to significantly change the nasal bioelectrics of the CF mice.


Subject(s)
Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis/therapy , Epithelial Cells/physiology , Genetic Therapy/methods , Nasal Mucosa/physiology , Promoter Regions, Genetic , Amiloride/pharmacology , Animals , Chloride Channels/metabolism , Cilia/physiology , Cystic Fibrosis/metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/analysis , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Gene Expression , Humans , Immunoprecipitation , Membrane Potentials , Mice , Mice, Knockout , Mice, Transgenic , Reverse Transcriptase Polymerase Chain Reaction , Sodium Channel Blockers/pharmacology , Trachea/physiology , Transgenes , Treatment Failure
11.
Am J Physiol Lung Cell Mol Physiol ; 290(2): L270-7, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16155086

ABSTRACT

The ion transport defects reported for human cystic fibrosis (CF) airways are reproduced in nasal epithelia of the CF mouse. Although this tissue has been studied in vivo using the nasal potential difference technique and as a native tissue mounted in the Ussing chamber, little information is available on cultured murine nasal epithelia. We have developed a polarized cell culture model of primary murine nasal epithelia in which the CF tissue exhibits not only a defect in cAMP-mediated Cl- secretion but also the Na+ hyperabsorption and upregulation of the Ca2+-activated Cl- conductance observed in human airways. Both the wild-type and CF cultures were constituted predominantly of undifferentiated cuboidal columnar cells, with most cultures exhibiting a small number of ciliated cells. Although no goblet cells were observed, RT-PCR demonstrated the expression of Muc5ac RNA after approximately 22 days in culture. The CF tissue exhibited an adherent layer of mucus similar to the mucus plaques reported in the distal airways of human CF patients. Furthermore, we found that treatment of CF preparations with a Na+ channel blocker for 7 days prevented formation of mucus adherent to epithelial surfaces. The cultured murine nasal epithelial preparation should be an excellent model tissue for gene transfer studies and pharmacological studies of Na+ channel blockers and mucolytic agents as well as for further characterization of CF ion transport defects. Culture of nasal epithelia from DeltaF508 mice will be particularly useful in testing drugs that allow DeltaF508 CFTR to traffic to the membrane.


Subject(s)
Cystic Fibrosis , Disease Models, Animal , Nasal Mucosa/cytology , Animals , Cell Culture Techniques/methods , Cell Differentiation , Cystic Fibrosis/physiopathology , Electrophysiology , Female , Male , Mice , Mucus/physiology , Nasal Mucosa/physiology , Sodium Channel Blockers/pharmacology
13.
Exp Lung Res ; 26(4): 257-71, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10923244

ABSTRACT

Using differential display, we have identified a novel gene, KPL1, induced in rat tracheal epithelial (RTE) cells grown under conditions which stimulate ciliogenesis. The KPL1 protein is predicted to contain a pleckstrin homology (PH) domain, which has been found in numerous signal transduction and cytoskeletal proteins. These domains are thought to function by recruiting proteins to cellular membranes, and they have been shown to bind phosphoinositols and the beta/gamma subunit of G proteins. We have cloned rat and human KPL1; the predicted protein translations are 94% identical. Alternate transcripts exist in rat and human tracheal cells that predict a protein which contains a 35-amino acid insert. KPL1 was upregulated in RTE cultures undergoing mucociliary but not squamous differentiation; and in cultures undergoing mucociliary differentiation, KPL1 expression most closely paralleled that of a marker of ciliated cell differentiation (axonemal dynein heavy chain) and not a marker of mucous cell differentiation (mucin 5AC). As a new member of the family of PH domain-containing proteins, KPL1 may have a unique role in ciliated cell differentiation or function.


Subject(s)
Cell Differentiation , Membrane Proteins , Proteins/genetics , Trachea/cytology , Amino Acid Sequence , Animals , Blood Proteins/chemistry , Cilia , Epithelial Cells/cytology , Humans , Intracellular Signaling Peptides and Proteins , Molecular Sequence Data , Phosphoproteins/chemistry , Rats , Rats, Inbred F344 , Sequence Homology, Amino Acid , Up-Regulation
14.
J Leukoc Biol ; 66(2): 310-4, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10449174

ABSTRACT

To further enhance chimerism, 229 primary allograft recipients have received perioperative intravenous infusion of a single dose of 3 to 6 X 10(8) unmodified donor bone marrow (BM) cells/kg body weight. In addition, 42 patients have been accrued in a concurrent protocol involving multiple (up to three) sequential perioperative infusions of 2 x 10(8) BM cells/kg/day from day 0-2 posttransplantation (PTx). Organ recipients (n = 133) for whom BM was not available were monitored as controls. The infusion of BM was safe and except for 50 (18%), all study patients have optimal graft function. Of the control patients, allografts in 30 (23%) have been lost during the course of follow-up. The cumulative risk of acute cellular rejection (ACR) was statistically lower in the study patients compared with that of controls. It is interesting that, 62% of BM-augmented heart recipients were free of ACR (Grade > or = 3A) in the first 6 months PTx compared to controls. The incidence of obliterative bronchiolitis was also statistically lower in study lung recipients (3.8%) compared with the contemporaneously acquired controls (31%). The levels of donor cell chimerism were at least a log higher in the peripheral blood of majority of the study patients compared with that of controls. The incidence of donor-specific hyporeactivity, as determined by one-way mixed leukocyte reaction, was also higher in those BM-augmented liver, kidney, and lung recipients that could be evaluated compared to controls.


Subject(s)
Bone Marrow Cells/immunology , Bone Marrow Transplantation/immunology , Immune Tolerance/immunology , Leukocyte Transfusion , Leukocytes/immunology , Organ Transplantation , Female , Follow-Up Studies , Graft Rejection/immunology , Heart Transplantation/immunology , Herpesvirus 4, Human/immunology , Humans , Incidence , Kidney Transplantation/immunology , Liver Transplantation/immunology , Lung Transplantation/immunology , Lymphoproliferative Disorders/epidemiology , Lymphoproliferative Disorders/immunology , Transplantation Chimera/immunology , Transplantation, Homologous/immunology
15.
J Cell Biol ; 145(6): 1277-92, 1999 Jun 14.
Article in English | MEDLINE | ID: mdl-10366599

ABSTRACT

It is well established that mutations in the presenilin 1 and 2 genes cause the majority of early onset Alzheimer's disease (AD). However, our understanding of the cellular functions of the proteins they encode remains rudimentary. Knowledge of proteins with which the presenilins interact should lead to a better understanding of presenilin function in normal and disease states. We report here the identification of a calcium-binding protein, calmyrin, that interacts preferentially with presenilin 2 (PS2). Calmyrin is myristoylated, membrane-associated, and colocalizes with PS2 when the two proteins are overexpressed in HeLa cells. Yeast two-hybrid liquid assays, affinity chromatography, and coimmunoprecipitation experiments confirm binding between PS2 and calmyrin. Functionally, calmyrin and PS2 increase cell death when cotransfected into HeLa cells. These results allude to several provocative possibilities for a dynamic role of calmyrin in signaling, cell death, and AD.


Subject(s)
Calcium-Binding Proteins/metabolism , Calcium/metabolism , Lipoproteins/metabolism , Membrane Proteins/metabolism , Myristic Acids/metabolism , Alzheimer Disease/genetics , Alzheimer Disease/metabolism , Amino Acid Sequence , Apoptosis , Calcium-Binding Proteins/analysis , Calcium-Binding Proteins/chemistry , Calcium-Binding Proteins/genetics , Cell Nucleus/metabolism , Chromatography, Affinity , Cytoplasm/metabolism , Endoplasmic Reticulum/metabolism , Fluorescent Antibody Technique , Gene Expression , HeLa Cells , Humans , Lipoproteins/analysis , Lipoproteins/chemistry , Lipoproteins/genetics , Membrane Proteins/analysis , Membrane Proteins/chemistry , Membrane Proteins/genetics , Molecular Sequence Data , Peptide Fragments/chemistry , Peptide Fragments/genetics , Peptide Fragments/metabolism , Precipitin Tests , Presenilin-2 , Protein Binding , RNA, Messenger/analysis , RNA, Messenger/genetics , RNA, Messenger/metabolism , Transfection , Yeasts/genetics , Yeasts/metabolism
16.
Am J Respir Cell Mol Biol ; 20(4): 675-83, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10100999

ABSTRACT

To identify genes upregulated during the process of ciliated cell differentiation of airway epithelial cells, differential display was used to compare RNA from rat tracheal epithelial (RTE) cells cultured under conditions that inhibit/promote ciliated cell differentiation. Several partial complementary DNAs (cDNAs) were identified whose expression was regulated coordinately with ciliated cell differentiation. One of these, KPL2, detected a messenger RNA transcript of approximately 6 kb when used as a probe on Northern blots of RNA from ciliated cultures but was undetectable in RNA from nonciliated cultures. Sequencing of overlapping clones obtained by a modified rapid amplification of cDNA ends procedure generated a complete cDNA sequence that exhibited no significant homology to sequences in GenBank, indicating that KPL2 is a novel gene. Southern analysis demonstrated that KPL2 exists as a single-copy gene. KPL2 contains a long open reading frame predicted to code for a protein of > 200 kD. Several putative functional motifs are present in the protein, including a calponin homology domain, three nuclear localization signals, a consensus P-loop, and a proline-rich region, suggesting that KPL2 has a unique function. KPL2 was undetectable in heart and liver samples, but was expressed in brain and testis, tissues that contain axonemal structures. In seminiferous tubules of the testis, KPL2 expression was stage-specific and appeared to be highest in spermatocytes and round spermatids. During differentiation of RTE cells, the expression of KPL2 closely paralleled that of an axonemal dynein heavy chain. These results suggest that KPL2 plays an important role in the differentiation or function of ciliated cells in the airway.


Subject(s)
Cilia/physiology , Epithelial Cells/physiology , Gene Expression Regulation , Microfilament Proteins/genetics , Trachea/cytology , Amino Acid Sequence , Animals , Cells, Cultured , Cloning, Molecular , Epidermal Growth Factor/pharmacology , Epithelial Cells/cytology , Epithelial Cells/drug effects , Male , Microfilament Proteins/biosynthesis , Microfilament Proteins/chemistry , Molecular Sequence Data , Organ Specificity , RNA, Messenger/genetics , Rats , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Reverse Transcriptase Polymerase Chain Reaction , Trachea/physiology , Transcription, Genetic , Tretinoin/pharmacology
18.
Am J Med Genet ; 82(2): 155-60, 1999 Jan 15.
Article in English | MEDLINE | ID: mdl-9934981

ABSTRACT

Primary ciliary dyskinesia (PCD) is a genetic disease characterized by abnormal ciliary structure and function, impaired mucociliary clearance, and chronic middle ear, sinus, and lung disease. PCD is associated with situs inversus in approximately 50% of the patients. One proposed explanation for this relationship is that normal ciliary function plays a role in normal organ orientation, whereas organ orientation in PCD is a random event because of dysfunctional cilia in early embryonic development. Another hypothesis for the association between PCD and situs inversus is that mutated genes in PCD not only cause defective cilia, but are also linked to the control of organ laterality, such that abnormalities in this molecular pathway result in random left-right asymmetry. We report on a set of monozygotic twin women with PCD. In both patients, deficiency of the inner dynein arms was noted on ciliary ultrastructural analysis, associated with a clinical syndrome of bronchiectasis, chronic sinusitis, and middle ear disease. One of the twins has situs solitus, the other has situs inversus totalis. DNA analysis confirmed that the twins are monozygotic. This is consistent with the hypothesis that situs inversus occurring in patients with primary ciliary dyskinesia is a random but "complete" event in the fetal development of patients with PCD.


Subject(s)
Ciliary Motility Disorders/physiopathology , Diseases in Twins , Situs Inversus/diagnostic imaging , Twins, Monozygotic , Adult , Cilia/ultrastructure , Female , Humans , Microscopy, Electron , Radiography , Situs Inversus/physiopathology
19.
Am J Physiol ; 273(5): L1082-9, 1997 11.
Article in English | MEDLINE | ID: mdl-9374738

ABSTRACT

Glutathione plays an essential role in protecting the pulmonary system from toxic insults. gamma-Glutamyl transpeptidase-related enzyme (GGT-rel) is a novel protein capable of cleaving the gamma-glutamyl peptide bond of glutathione and of converting leukotriene C4 to leukotriene D4. A rat homologue of GGT-rel was identified and was found to be highly expressed in cultures of differentiating rat tracheal epithelial (RTE) cells. The 2.6-kb cDNA predicts a 572-amino acid protein with 79% identity to human GGT-rel. GGT-rel was weakly expressed in normal trachea but was strongly induced by epidermal growth factor in cultures of RTE cells. GGT-rel was also highly expressed in lung tumors induced by inhalation of isobutyl nitrite. These results demonstrate that GGT-rel 1) is expressed in normal tracheal cells, 2) can be induced by epidermal growth factor, and 3) is elevated after chemical exposure. The induction of high levels of GGT-rel may play an important role in protecting the lung from oxidative stress or other toxic insults.


Subject(s)
Gene Expression Regulation, Enzymologic , Lung Neoplasms/enzymology , Trachea/enzymology , gamma-Glutamyltransferase/biosynthesis , Amino Acid Sequence , Animals , Base Sequence , Carcinogens , Cell Differentiation/drug effects , Cells, Cultured , DNA Primers , Epidermal Growth Factor/pharmacology , Epithelial Cells/cytology , Epithelial Cells/enzymology , Gene Expression Regulation, Enzymologic/drug effects , Humans , Lung Neoplasms/chemically induced , Molecular Sequence Data , Nitrites , Organ Specificity , Polymerase Chain Reaction , Rats , Sequence Alignment , Sequence Homology, Amino Acid , Trachea/cytology , gamma-Glutamyltransferase/chemistry
20.
In Vitro Cell Dev Biol Anim ; 33(3): 212-7, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9112131

ABSTRACT

We are studying the regulation of ciliated cell differentiation using an in vitro model of tracheal regeneration. Previously, we reported that removal of growth stimulating compounds such as epidermal growth factor (EGF) and cholera toxin reduced DNA synthesis and cell number while increasing ciliated cell differentiation (Clark et al., 1995). This result suggested that the induction of growth arrest may stimulate terminal differentiation of airway epithelial cells into ciliated cells. Transforming growth factor beta s (TGF beta s) inhibit epithelial cell proliferation and have also been shown to stimulate epithelial cell differentiation. In this study, the effect of TGF beta 1 on growth and ciliated cell differentiation of rat tracheal epithelial (RTE) cells was examined. TGF beta 1 inhibited [3H]thymidine incorporation by RTE cells in a dose-dependent manner. A 40% inhibition was observed after a 24-h incubation with 10 pM TGF beta 1. Continuous treatment with TGF beta 1 (1-50 pM) also reduced cell number during the time when ciliogenesis occurs. This reduction resulted in part from a loss of cells through exfoliation, in addition to the inhibition of proliferation. The exfoliated cells exhibited several morphological features characteristic of apoptosis, including shrunken cells, condensed and fragmented nuclei, and intact organelles. In addition, electrophoretic analysis of genomic DNA analysis isolated from exfoliated cells demonstrated the presence of a nucleosomal ladder. However, in contrast to the removal of EGF1 treatment with TGF beta 1 for 7 d did not increase ciliated cell differentiation. TGF beta 1 is, therefore, capable of inhibiting proliferation and increasing apoptosis in RTE cells without stimulating ciliated cell differentiation.


Subject(s)
Apoptosis , Cell Differentiation , Cell Division , Trachea/cytology , Transforming Growth Factor beta/pharmacology , Animals , Cell Count , Cells, Cultured , Cilia , DNA/biosynthesis , Epithelial Cells , Male , Microscopy, Electron , Rats , Rats, Inbred F344
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