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1.
Int J Biol Macromol ; 272(Pt 2): 132883, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38838898

ABSTRACT

Glycyrrhiza glabra extract is widely known for its antioxidant and anti-inflammatory properties and can improve the wound healing process. The aim of this work was to shorten the time of the healing process by using an eco-sustainable wound dressing based on Spanish broom flexible cellulosic fabric by impregnation with G. glabra extract-loaded ethosomes. Chemical analysis of G. glabra extract was performed by LC-DAD-MS/MS and its encapsulation into ethosomes was obtained using the ethanol injection method. Lipid vesicles were characterized in terms of size, polydispersity index, entrapment efficiency, zeta potential, and stability. In vitro release studies, biocompatibility, and scratch test on 3T3 fibroblasts were performed. Moreover, the structure of Spanish broom dressing and its ability to absorb wound exudate was characterized by Synchrotron X-ray phase contrast microtomography (SR-PCmicroCT). Ethosomes showed a good entrapment efficiency, nanometric size, good stability over time and a slow release of polyphenols compared to the free extract, and were not cytotoxic. Lastly, the results revealed that Spanish broom wound dressing loaded with G. glabra ethosomes is able to accelerate wound closure by reducing wound healing time. To sum up, Spanish broom wound dressing could be a potential new green tool for biomedical applications.


Subject(s)
Bandages , Cellulose , Glycyrrhiza , Plant Extracts , Spartium , Wound Healing , Animals , Mice , Glycyrrhiza/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Wound Healing/drug effects , Cellulose/chemistry , Cellulose/pharmacology , Spartium/chemistry , 3T3 Cells
2.
Hernia ; 22(5): 827-836, 2018 10.
Article in English | MEDLINE | ID: mdl-29700716

ABSTRACT

PURPOSE: To compare the perioperative outcomes of initial, consecutive robotic-assisted transabdominal preperitoneal (R-TAPP) inguinal hernia repair (IHR) cases with consecutive open cases completed by the same surgeons. METHODS: Multicenter, retrospective, comparative study of perioperative results from open and robotic IHR using standard univariate and multivariate regression analyses for propensity score matched (1:1) cohorts. RESULTS: Seven general surgeons at six institutions contributed 602 consecutive open IHR and 652 consecutive R-TAPP IHR cases. Baseline patient characteristics in the unmatched groups were similar with the exception of previous abdominal surgery and all baseline characteristics were comparable in the matched cohorts. In matched analyses, postoperative complications prior to discharge were comparable. However, from post discharge through 30 days, fewer patients experienced complications in the R-TAPP group than in the open group [4.3% vs 7.7% (p = 0.047)]. The R-TAPP group had no reoperations post discharge through 30 days of follow-up compared with five patients (1.1%) in the open group (p = 0.062), respectively. Multivariate logistic regression analysis which demonstrated patient age > 65 years and the open approach were risk factors for complications within 30 days post discharge in the matched group [age > 65 years: odds ratio (OR) = 3.33 (95% CI 1.89, 5.87; p < 0.0001); open approach: OR = 1.89 (95% CI 1.05, 3.38; p = 0.031)]. CONCLUSIONS: In this matched analysis, R-TAPP provides similar postoperative complications prior to discharge and a lower rate of postoperative complications through 30 days compared to open repair. R-TAPP is a promising and reproducible approach, and may facilitate adoption of minimally invasive repairs of inguinal hernias.


Subject(s)
Hernia, Inguinal/surgery , Herniorrhaphy/methods , Laparoscopy , Robotic Surgical Procedures , Age Factors , Female , Herniorrhaphy/adverse effects , Humans , Male , Matched-Pair Analysis , Middle Aged , Postoperative Complications , Retrospective Studies , Risk Factors
3.
Lab Chip ; 17(7): 1340-1348, 2017 03 29.
Article in English | MEDLINE | ID: mdl-28276545

ABSTRACT

A microfluidic system that combines membraneless microfluidic dialysis and dielectrophoresis to achieve label-free isolation and concentration of bacteria from whole blood is presented. Target bacteria and undesired blood cells are discriminated on the basis of their differential susceptibility to permeabilizing agents that alter the dielectrophoretic behavior of blood cells but not bacteria. The combined membraneless microdialysis and dielectrophoresis system isolated 79 ± 3% of Escherichia coli and 78 ± 2% of Staphylococcus aureus spiked into whole blood at a processing rate of 0.6 mL h-1. Collection efficiency was independent of the number of target bacteria up to 105 cells. Quantitative PCR analysis revealed that bacterial 16S rDNA levels were enriched more than 307-fold over human DNA in the fraction recovered from the isolation system compared with the original specimen. These data demonstrate feasibility for an instrument to accelerate the detection and analysis of bacteria in blood by first isolating and concentrating them in a microchamber.


Subject(s)
Bacteremia , Bacteria/isolation & purification , Bacterial Load/methods , Electrophoresis/instrumentation , Microfluidic Analytical Techniques/instrumentation , Bacteremia/diagnosis , Bacteremia/microbiology , Bacteria/genetics , Computer Simulation , DNA, Bacterial/analysis , Equipment Design , Humans , RNA, Ribosomal, 16S/genetics
5.
EXCLI J ; 15: 166-76, 2016.
Article in English | MEDLINE | ID: mdl-27103896

ABSTRACT

Depressed levels of atheroprotective large HDL particles are common in obesity and cardiovascular disease (CVD). Increases in large HDL particles are favourably associated with reduced CVD event risk and coronary plaque burden. The objective of the study is to compare the effectiveness of low-carbohydrate diets and weight loss for increasing blood levels of large HDL particles at 1 year. This study was performed by screening for body mass index (BMI) and metabolic syndrome in 160 consecutive subjects referred to our out-patient Metabolic Unit in South Italy. We administered dietary advice to four small groups rather than individually. A single team comprised of a dietitian and physician administered diet-specific advice to each group. Large HDL particles at baseline and 1 year were measured using two-dimensional gel electrophoresis. Dietary intake was assessed via 3-day diet records. Although 1-year weight loss did not differ between diet groups (mean 4.4 %), increases in large HDL particles paralleled the degree of carbohydrate restriction across the four diets (p<0.001 for trend). Regression analysis indicated that magnitude of carbohydrate restriction (percentage of calories as carbohydrate at 1 year) and weight loss were each independent predictors of 1-year increases in large HDL concentration. Changes in HDL cholesterol concentration were modestly correlated with changes in large HDL particle concentration (r=0.47, p=.001). In conclusion, reduction of excess dietary carbohydrate and body weight improved large HDL levels. Comparison trials with cardiovascular outcomes are needed to more fully evaluate these findings.

6.
Osteoporos Int ; 26(12): 2785-91, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26068297

ABSTRACT

UNLABELLED: We evaluated the effect of parathyroid hormone (PTH) on Wnt10b production by immune system cells in humans. We showed that bone anabolic effect of intermittent PTH treatment may be amplified by T cells through increased production of Wnt10b. Chronic increase in PTH as in primary hyperparathyroidism does not increase Wnt10b expression. INTRODUCTION: The aim of this study is to assess the effect of PTH on Wnt10b production by immune system cells in humans. We assessed both the effect of intermittent PTH administration (iPTH) and of chronic PTH hypersecretion in primary hyperparathyroidism (PHP). METHODS: Eighty-two women affected by post-menopausal osteoporosis were randomly assigned to treatment with calcium and vitamin D alone (22) or plus 1-84 PTH (42), or intravenous ibandronate (18). Wnt10b production by unfractioned blood nucleated cells and by T, B cells and monocytes was assessed by real-time RT-PCR and ELISA at baseline, 3, 6, 12 and 18 months of treatment. The effect of chronic elevation of PTH was evaluated in 20 patients affected by PHP at diagnosis and after surgical removal of parathyroid adenoma. WNT10b from both osteoporotic and PHP patients was compared to healthy subjects matched for age and sex. RESULTS: iPTH increases Wnt10b production by T cells, whereas PHP does not. After surgical restoration of normal parathyroid function, WNT10b decreases, although it is still comparable with healthy subjects' level. Thus, chronic elevation of PTH does not significantly increase WNT10b production as respect to control. CONCLUSIONS: This is the first work showing the effect of both intermittent and chronic PTH increase on Wnt10b production by immune system cells. We suggest that, in humans, T cells amplified the anabolic effect of PTH on bone, by increasing Wnt10b production, which stimulates osteoblast activity.


Subject(s)
Osteoporosis, Postmenopausal/drug therapy , Parathyroid Hormone/therapeutic use , Proto-Oncogene Proteins/biosynthesis , T-Lymphocytes/metabolism , Wnt Proteins/biosynthesis , Aged , Aged, 80 and over , Bone Density Conservation Agents/administration & dosage , Bone Density Conservation Agents/therapeutic use , Calcium/therapeutic use , Diphosphonates/administration & dosage , Diphosphonates/therapeutic use , Drug Administration Schedule , Drug Therapy, Combination , Female , Humans , Hyperparathyroidism, Primary/blood , Ibandronic Acid , Middle Aged , Osteoporosis, Postmenopausal/blood , Osteoporosis, Postmenopausal/immunology , Parathyroid Hormone/administration & dosage , Parathyroid Hormone/blood , Proto-Oncogene Proteins/genetics , RNA, Messenger/genetics , Vitamin D/therapeutic use , Wnt Proteins/genetics
7.
Vet Comp Orthop Traumatol ; 26(4): 271-9, 2013.
Article in English | MEDLINE | ID: mdl-23857571

ABSTRACT

OBJECTIVE: To evaluate the relative position of the femur and tibia in cranial cruciate ligament-intact stifles, cranial cruciate ligament-deficient stifles, and cruciate-deficient stifles following lateral extracapsular suture system (LESS) stabilization under load at specific joint angles. STUDY DESIGN: In vitro biomechanical study. METHODS: Twenty pelvic limbs from 11 dogs were used to evaluate the relative position of the femur and tibia between three stifle conditions (cranial cruciate ligament-intact, cranial cruciate ligament-deficient, and LESS-stabilized) at a load of 30% of body weight and stifle angles of 125°, 135°, and 145° using electromagnetic tracking sensors. RESULTS: Cranial cruciate ligament-deficient stifles had significantly greater (p <0.0001) cranial displacement and internal rotation of the tibia relative to the femur than cranial cruciate ligament-intact or LESS stifles at all stifle angles. Cranial displacement of the tibia relative to the femur for cranial cruciate ligament-intact and LESS stifles were not significantly different from one another at a stifle angle of 125°, but were significantly different at stifle angles of 135° (p = 0.0182) and 145° (p = 0.0012). There was no significant difference in internal rotation of the tibia relative to the femur between the cranial cruciate ligament-intact and LESS stifles at any of the stifle angles. CLINICAL SIGNIFICANCE: The LESS procedure effectively decreased cranial tibial displacement and eliminated internal rotation of the tibia relative to the femur in the cranial cruciate ligament-deficient stifles at stifle angles of 125°, 135°, and 145° in vitro.


Subject(s)
Anterior Cruciate Ligament/surgery , Dogs , Stifle/surgery , Suture Techniques/veterinary , Animals , Biomechanical Phenomena , Bone Screws/veterinary , Cadaver
8.
Br J Cancer ; 108(12): 2525-36, 2013 Jun 25.
Article in English | MEDLINE | ID: mdl-23801032

ABSTRACT

BACKGROUND: Bone metastases represent a common and severe complication in breast cancer, and the involvement of cancer stem cells (CSCs) in the promotion of bone metastasis is currently under discussion. Here, we used a human-in-mice model to study bone metastasis formation due to primary breast CSCs-like colonisation. METHODS: Primary CD44⁺CD24⁻ breast CSCs-like were transduced by a luciferase-lentiviral vector and injected through subcutaneous and intracardiac (IC) routes in non-obese/severe-combined immunodeficient (NOD/SCID) mice carrying subcutaneous human bone implants. The CSCs-like localisation was monitored by in vivo luciferase imaging. Bone metastatic CSCs-like were analysed through immunohistochemistry and flow cytometry, and gene expression analyses were performed by microarray techniques. RESULTS: Breast CSCs-like colonised the human-implanted bone, resulting in bone remodelling. Bone metastatic lesions were histologically apparent by tumour cell expression of epithelial markers and vimentin. The bone-isolated CSCs-like were CD44⁻CD24⁺ and showed tumorigenic abilities after injection in secondary mice. CD44⁻CD24⁺ CSCs-like displayed a distinct bone tropism signature that was enriched in genes that discriminate bone metastases of breast cancer from metastases at other organs. CONCLUSION: Breast CSCs-like promote bone metastasis and display a CSCs-like bone tropism signature. This signature has clinical prognostic relevance, because it efficiently discriminates osteotropic breast cancers from tumour metastases at other sites.


Subject(s)
Bone Neoplasms/secondary , Bone and Bones/metabolism , Breast Neoplasms/pathology , Carcinoma/pathology , Neoplastic Stem Cells/pathology , Transcriptome , Adult , Animals , Bone Neoplasms/genetics , Bone and Bones/pathology , Breast Neoplasms/genetics , Carcinoma/genetics , Female , Gene Expression Regulation, Neoplastic , Genes, Switch/genetics , Humans , Mice , Mice, Inbred NOD , Mice, SCID , Neoplastic Stem Cells/metabolism , Organ Specificity/genetics , Phenotype , Transcriptome/physiology
9.
Oncogenesis ; 2: e43, 2013 Apr 08.
Article in English | MEDLINE | ID: mdl-23567620

ABSTRACT

In non-small cell lung cancer (NSCLC), receptor tyrosine kinases (RTKs) stand out among causal dominant oncogenes, and the ablation of RTK signaling has emerged as a novel tailored therapeutic strategy. Nonetheless, long-term RTK inhibition leads invariably to acquired resistance, tumor recurrence and metastatic dissemination. In ALK+ cell lines, inhibition of ALK signaling was associated with coactivation of several RTKs, whose pharmacological suppression reverted the partial resistance to ALK blockade. Remarkably, ERBB2 signaling synergized with ALK and contributed to the neoplastic phenotype. Moreover, the engagement of wild-type epidermal growth factor receptor or MET receptors could sustain cell viability through early growth response 1 (EGR1) and/or Erk1/2; Akt activation and EGR1 overexpression prevented cell death induced by combined ALK/RTK inhibition. Membrane expression of ERBB2 in a subset of primary naive ALK+ NSCLC could be relevant in the clinical arena. Our data demonstrate that the neoplastic phenotype of ALK-driven NSCLC relays 'ab initio' on the concomitant activation of multiple RTK signals via autocrine/paracrine regulatory loops. These findings suggest that molecular and functional signatures are required in de novo lung cancer patients for the design of efficacious and multi-targeted 'patient-specific' therapies.

10.
Vet Comp Orthop Traumatol ; 26(3): 208-17, 2013.
Article in English | MEDLINE | ID: mdl-23460337

ABSTRACT

OBJECTIVE: To evaluate the effect of two tibial attachment sites for lateral suture stabilization (LSS) on the three-dimensional femorotibial translational and rotational movements of the cranial cruciate ligament- deficient canine stifle during the early, middle and late stance phases. STUDY DESIGN: In vitro biomechanical study: 32 hindlimbs from 16 canine cadavers. METHODS: Limbs were mounted in a testing jig and an electromagnetic tracking system was used to determine the three-dimensional femorotibial translational and rotational movements under 33% of body weight load during early, middle and late stance in the following sequence: cranial cruciate ligament-intact, cranial cruciate ligament-deficient and LSS with the distal anchor through the tibial tuberosity (LSSTT) or through the cranial eminence of the extensor groove (LSSEG). The proximal anchor point was the lateral femorofabellar ligament. RESULTS: Post-LSS stifle three-dimensional femorotibial translational and rotational movements were more comparable to normal than post-transection movements for both techniques. Both LSS techniques restored femorotibial movements in cranial cruciate ligament-deficient stifles to varying amounts but neither technique successfully restored normal three-dimensional femorotibial movements. The LSSEG improved femorotibial movements of the cranial cruciate ligament-deficient stifle in the medial-lateral direction and axial rotation but performed poorly in restoring femorotibial movements in the cranial-caudal direction as compared to the LSSTT. CLINICAL SIGNIFICANCE: Both the LSSTT and LSSEG techniques failed to completely restore normal three-dimensional femorotibial translational and rotational movements in cranial cruciate ligament-deficient stifles in vitro.


Subject(s)
Anterior Cruciate Ligament/surgery , Dogs , Stifle/surgery , Suture Techniques/veterinary , Animals , Biomechanical Phenomena , Cadaver
11.
Osteoporos Int ; 23(4): 1245-53, 2012 Apr.
Article in English | MEDLINE | ID: mdl-21617993

ABSTRACT

UNLABELLED: This study shows that teriparatide promotes the circulating osteoblast (OB) precursor degree of maturation in patients affected by postmenopausal osteoporosis. INTRODUCTION: Anabolic treatment with teriparatide has proven effective for the therapy of postmenopausal osteoporosis and significantly reduces the risk of non-vertebral fragility fractures. The aim of this study was to investigate the effect of teriparatide on circulating OB precursors. METHODS: We evaluated by flow cytometry and real-time PCR the expression of OBs typical markers in peripheral blood mononuclear cells during treatment with teriparatide plus calcium and vitamin D, raloxifene plus calcium and vitamin D or calcium and vitamin D alone at various time points. Serum bone alkaline phosphatase and osteocalcin (OC) were measured as markers of bone turnover. RESULTS: Our results show that circulating OB precursors are more numerous and more immature in patients affected by fragility fractures than in osteoporotic patients without fractures. We also show that teriparatide treatment increases the expression of alkaline phosphatase and of OC in OB precursors; thus, it increases their degree of maturation. CONCLUSIONS: We suggest that teriparatide acts as anabolic agents also by promoting the maturation of OB precursors.


Subject(s)
Bone Density Conservation Agents/pharmacology , Osteoblasts/drug effects , Osteoporosis, Postmenopausal/blood , Teriparatide/pharmacology , Aged , Alkaline Phosphatase/blood , Biomarkers/blood , Bone Density Conservation Agents/therapeutic use , Calcium/pharmacology , Calcium/therapeutic use , Cell Differentiation/drug effects , Drug Therapy, Combination , Female , Humans , Mesenchymal Stem Cells/drug effects , Middle Aged , Osteoblasts/pathology , Osteocalcin/blood , Osteoporosis, Postmenopausal/complications , Osteoporosis, Postmenopausal/drug therapy , Osteoporotic Fractures/blood , Osteoporotic Fractures/etiology , Osteoporotic Fractures/prevention & control , Raloxifene Hydrochloride/pharmacology , Raloxifene Hydrochloride/therapeutic use , Secondary Prevention , Teriparatide/therapeutic use , Vitamin D/pharmacology , Vitamin D/therapeutic use
12.
Osteoporos Int ; 22(11): 2869-77, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21116815

ABSTRACT

UNLABELLED: This study evaluates cytokines production in bone and bone marrow of patients with an osteoporotic fracture or with osteoarthritis by real time PCR, Western blot and immunohistochemistry. We demonstrate that the cytokine pattern is shifted towards osteoclast activation and osteoblast inhibition in patients with osteoporotic fractures. INTRODUCTION: Fragility fractures are the resultant of low bone mass and poor bone architecture typical of osteoporosis. Cytokines involved in the control of bone cell maturation and function are produced by both bone itself and bone marrow cells, but the roles of these two sources in its control and the amounts they produce are not clear. This study compares their production in patients with an osteoporotic fracture and those with osteoarthritis. METHODS: We evaluated 52 femoral heads from women subjected to hip-joint replacement surgery for femoral neck fractures due to low-energy trauma (37), or for osteoarthritis (15). Total RNA was extracted from both bone and bone marrow, and quantitative PCR was used to identify the receptor activator of nuclear factor kB Ligand (RANKL), osteoprotegerin (OPG), macrophage colony stimulating factor (M-CSF), transforming growth factor ß (TGFß), Dickoppf-1 (DKK-1) and sclerostin (SOST) expression. Immunohistochemistry and Western blot were performed in order to quantify and localize in bone and bone marrow the cytokines. RESULTS: We found an increase of RANKL/OPG ratio, M-CSF, SOST and DKK-1 in fractured patients, whereas TGFß was increased in osteoarthritic bone. Bone marrow produced greater amounts of RANKL, M-CSF and TGFß compared to bone, whereas the production of DKK-1 and SOST was higher in bone. CONCLUSIONS: We show that bone marrow cells produced the greater amount of pro-osteoclastogenic cytokines, whereas bone cells produced higher amount of osteoblast inhibitors in patients with fragility fracture, thus the cytokine pattern is shifted towards osteoclast activation and osteoblast inhibition in these patients.


Subject(s)
Bone Marrow/metabolism , Cytokines/metabolism , Femur Head/metabolism , Osteoarthritis/metabolism , Osteoporotic Fractures/metabolism , Adaptor Proteins, Signal Transducing , Aged , Aged, 80 and over , Blotting, Western , Bone Morphogenetic Proteins/metabolism , Female , Genetic Markers , Humans , Intercellular Signaling Peptides and Proteins/metabolism , Macrophage Colony-Stimulating Factor/metabolism , Middle Aged , Osteoblasts/metabolism , Osteoclasts/metabolism , Osteoprotegerin/metabolism , RANK Ligand/metabolism , Real-Time Polymerase Chain Reaction , Transforming Growth Factor beta/metabolism
13.
J Microsc ; 241(2): 207-11, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21118220

ABSTRACT

To understand the reported difference between double band, sarcomeric second harmonic generation pattern of isolated myofibril and predominant single band pattern found in thick muscle tissues, we studied the effect of myofibril preparation on the second harmonic generation pattern. We found that double band sarcomeric second harmonic generation pattern usually observed in myofibrils (prepared from fresh tissue) is due to muscle alteration during the mixing and triton treatment processes. Single band sarcomeric second harmonic generation pattern could be observed in isolated myofibrils when this alteration is previously prevented using paraformaldehyd fixed tissue. We conclude that single band sarcomeric second harmonic generation pattern is a signature of adult muscle myofibrils in normal physiological condition, suggesting that sarcomeric second harmonic generation patterns could be used as a valuable diagnosis tool of muscle health.


Subject(s)
Muscle, Skeletal/physiology , Muscle, Skeletal/ultrastructure , Myofibrils/ultrastructure , Sarcomeres/ultrastructure , Animals , Microscopy, Confocal , Models, Biological , Specimen Handling/methods , Tissue Preservation/methods , Xenopus laevis
14.
Plant Dis ; 89(1): 107, 2005 Jan.
Article in English | MEDLINE | ID: mdl-30795299

ABSTRACT

Inonotus rickii (Pat.) D. Reid (Hymenochaetaceae), widespread in tropical and subtropical countries, produces cankers and decay in several hardwoods. In Italy, it was recorded for the first time approximately 20 years ago on Parkinsonia sp. and Schinus molle L. in Sicily (2) and more recently on Acer negundo L. and Celtis australis L. in Rome (1). During May 2004, a survey was performed in Sicily in public and botanical gardens where the fungus had been detected in the past. Although the previously attacked plants had been removed, more trees were showing decline symptoms, with sparse foliage and some dieback. Typical brown, powdery masses of chlamydospores of I. rickii were observed near wounds or pruning cuts on Quercus cerris L. in Palermo and Sambucus nigra L. and Aberia caffra Hook. F. & Harv. in Catania. From each fungal specimen, fragments were isolated and cultured on malt agar. Cultures produced acute, thick-walled, brown setae (as much as 250 µm long) and abundant chlamydospores that were yellowish brown, thick walled, irregularly shaped, and 10 to 12 × 8 to 12 µm. Each isolate conformed to I. rickii (3). Cultures and voucher specimens have been deposited in the ISPaVe collection (PF40, PF41, and PF42) and the ROPV herbarium (772, 773, and 774). To our knowledge, this is the first report of I. rickii in living Q. cerris and A. caffra. S. nigra is a new host in Italy. This record shows that the pathogen has fully established itself in southern Europe where it may expand its host range. References: (1) T. Annesi et al. For. Pathol. 33:405, 2003. (2) M. Intini. Micol. Ital. 20:49, 1988. (3) J. A. Stalpers. Karstenia 40:167, 2000.

15.
Dev Dyn ; 222(4): 611-24, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11748830

ABSTRACT

The yolk syncytial layer (YSL) of the teleostean yolk cell is known to play important roles in the induction of cellular mesendoderm, as well as the patterning of dorsal tissues. To determine how this extraembryonic endodermal compartment is subdivided and morphologically transformed during early development, we have examined collective movements of vitally stained YSL nuclei in axiating zebrafish embryos by using four-dimensional confocal microscopy. During blastulation, gastrulation, and early segmentation, zebrafish YSL nuclei display several highly patterned movements, which are organized into spatially distinct morphogenetic domains along the anterior-posterior and dorsal-ventral axes. During the late blastula period, with the onset of epiboly, nuclei throughout the YSL initiate longitudinal movements that are directed along the animal-vegetal axis. As epiboly progresses, nuclei progressively recede from the advancing margin of the epibolic YSL. However, a small group of nuclei is retained at the YSL margin to form a constricting blastoporal ring. During mid-gastrulation, YSL nuclei undergo convergent-extension behavior toward the dorsal midline, with a subset of nuclei forming an axial domain that underlies the notochord. These highly patterned movements of YSL nuclei share remarkable similarities to the morphogenetic movements of deep cells in the overlying zebrafish blastoderm. The macroscopic shape changes of the zebrafish yolk cell, as well as the morphogenetic movements of its YSL nuclei, are homologous to several morphogenetic behaviors that are regionally expressed within the vegetal endodermal cell mass of gastrulating Xenopus embryos. In contrast to the cellular endoderm of Xenopus, the dynamics of zebrafish YSL show that a syncytial endodermal germ layer can express a temporal sequence of morphogenetic domains without undergoing progressive steps of cell fate restriction.


Subject(s)
Giant Cells/physiology , Zebrafish/embryology , Animals , Blastocyst/physiology , Cell Movement , Cell Nucleus/physiology , Embryo, Nonmammalian/physiology , Gastrula/physiology , Giant Cells/cytology
16.
J Allergy Clin Immunol ; 108(5): 703-8, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11692092

ABSTRACT

BACKGROUND: Recently, several studies have shown that heparin possesses various anti-inflammatory and antiallergic properties. It has been proposed that heparin might play an important role in limiting the inflammatory events associated with asthma and allergic rhinitis by neutralizing inflammatory mediators, such as eosinophil cationic protein and major basic protein, and by limiting eosinophil recruitment. OBJECTIVE: To test the hypothesis that heparin can limit the extent and magnitude of eosinophilic inflammation, we examined the effect of inhaled intranasal heparin on nasal response to allergen challenge in 10 patients with allergic rhinitis. METHODS: The capacity of heparin to reduce nasal response was studied by evaluating symptom score, eosinophil cationic protein concentration, and eosinophil counts in nasal lavage fluids 10, 60, and 360 minutes after allergen challenge. RESULTS: Pretreatment with intranasal heparin produced a significant reduction in symptom score 10 minutes after allergen challenge and reduced the eosinophil influx at each time point after antigen challenge, statistical significance being reached 60 and 360 minutes after allergen challenge. Similarly, the amount of eosinophil cationic protein in the nasal wash was reduced at each time point; this reached statistical significance 360 minutes after allergic challenge. CONCLUSION: Heparin was shown to provide protection with respect to nasal allergen challenge. The mechanism by which heparin produces its protective activity seems to be related to the neutralization of eosinophil cationic protein as well as to the reduction of eosinophil recruitment.


Subject(s)
Eosinophilia/drug therapy , Heparin/therapeutic use , Rhinitis, Allergic, Seasonal/drug therapy , Ribonucleases , Administration, Intranasal , Adolescent , Adult , Allergens/immunology , Asthma/drug therapy , Blood Proteins/biosynthesis , Cross-Over Studies , Double-Blind Method , Eosinophil Granule Proteins , Eosinophilia/immunology , Female , Heparin/administration & dosage , Humans , Kinetics , Leukocyte Count , Male , Nasal Lavage Fluid/immunology , Rhinitis, Allergic, Seasonal/diagnosis , Rhinitis, Allergic, Seasonal/immunology
17.
Am J Respir Cell Mol Biol ; 25(5): 592-9, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11713101

ABSTRACT

The ability of lung fibroblasts to modulate the immune response has been evaluated by analyzing the synthesis and release of interleukin (IL)-10 and IL-12 by lipopolysaccharide (LPS)-stimulated peripheral blood monocytes exposed to pulmonary fibroblast conditioned medium (FCM). IL-10 and IL-12 contents and gene expression were markedly modified by treatment with FCM as measured by ELISA (+97.5 +/- 12.8% and -68 +/- 7.3% for IL-10 and IL-12, respectively), immunocytochemistry, and reverse transcriptase-polymerase chain reaction (RT-PCR). These effects appeared to be mediated by prostaglandin E(2) (PGE(2)) as the modified release of both cytokines was reduced by treatment with indomethacin and mimicked by addition of exogenous PGE(2.) As a result of the enhanced production of IL-10, exposure of LPS/interferon (IFN)-gamma-activated monocytes to FCM was also able to reduce the expression of the class II major histocompatibility complex (MHC) molecule, human leukocyte-associated antigen-DR (HLA-DR) (-51.8 +/- 8.7%) and of the costimulatory molecule, CD40 (-53.9 +/- 11.7%). The expression of both molecules was completely restored when monocytes were pretreated with a neutralizing anti-IL-10 monoclonal antibody. The FCM obtained from fibrotic lung fibroblasts was instead less efficacious in potentiating LPS-stimulated IL-10 release and, consequently, in reducing HLA-DR and CD40 expression, suggesting that an impairment of the immune regulation operated by fibroblasts may be involved in the maintenance of chronic pulmonary inflammation.


Subject(s)
Fibroblasts/immunology , Interleukin-10/genetics , Interleukin-12/genetics , Lung/cytology , Monocytes/immunology , Pneumonia/immunology , Adult , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , CD40 Antigens/analysis , CD40 Antigens/biosynthesis , Cell Communication/immunology , Cells, Cultured , Chronic Disease , Dinoprostone/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Flow Cytometry , Gene Expression/immunology , HLA-DR Antigens/analysis , HLA-DR Antigens/biosynthesis , Humans , Immunohistochemistry , Indomethacin/pharmacology , Interleukin-10/analysis , Interleukin-12/analysis , Lipopolysaccharides/pharmacology , Lung/immunology , Monocytes/cytology , Monocytes/drug effects , Pneumonia/pathology , Pulmonary Fibrosis/immunology , Pulmonary Fibrosis/pathology , RNA, Messenger/analysis
18.
Proc Biol Sci ; 268(1476): 1589-93, 2001 Aug 07.
Article in English | MEDLINE | ID: mdl-11487406

ABSTRACT

The evolution of body size is a dominant feature of animal evolution. However, little is known about how the underlying developmental mechanisms that determine size change as body size evolves. Here we report on a case of body size evolution in the tobacco hornworm Manduca sexta that occurred over a period of nearly 30 years. We take advantage of an extensive series of physiological studies performed in the early 1970s that established the parameters that regulate body size in this species and compare their values with those of modern individuals that are descendants of the same colony. We show that three of the five processes that determine adult body size changed during this period, while two remained constant. Changes in these three developmental processes completely account for the observed evolutionary change in body size.


Subject(s)
Manduca/physiology , Animals , Biological Evolution , Body Weight/physiology
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