bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histomorphometric assessment of Haversian canal and osteocyte lacunae in different‐sized osteons in human rib
Authors: Qiu Shijing , Fyhrie David P. , Palnitkar Saroj , Rao D. Sudhaker 
Published: May 2003

AbstractThere is no detailed information available concerning the variations in bone, the Haversian canal, and osteocyte populations in different‐sized osteons. In this study a total of 398 secondary osteons were measured in archived rib sections from nine white men (20–25 years old). The sections were stained with basic fuchsin. The parameters included the osteon area (On.Ar), Haversian canal area (HC.Ar) and perimeter (HC.Pm), bone area (B.Ar), and osteocyte lacunar number (Lc.N). From these primary measurements the following indices were deduced: 1) lacunar number per bone area (Lc.N/B.Ar) and per osteon (Lc.N/On); 2) the ratio between Haversian canal perimeter and bone area (HC.Pm/B.Ar); and 3) the fraction of Haversian canal area (HC.Ar/On.Ar) and its complement, the fraction of bone area (B.Ar/On.Ar). The results showed that the osteons varied greatly in size, but very little in the fraction of bone area. Regression analyses showed that HC.Ar, HC.Pm, and Lc.N/On were positively associated with On.Ar (P < 0.001 for all). A significant negative correlation was found between On.Ar and Lc.N/B.Ar (P < 0.05) and HC.Pm/B.Ar (P < 0.0001). HC.Ar and HC.Pm increased significantly with increasing Lc.N/On (both P < 0.0001) rather than Lc.N/B.Ar. Lc.N/B.Ar had a significant positive correlation with HC.Ar/On.Ar (P < 0.05) and HC.Pm/B.Ar (P < 0.01). We conclude that: 1) the size of the osteon is determined by the quantum of bone removed by osteoclasts, 2) the osteon is well designed for molecular exchange, and 3) a well designed osteon may be produced via the regulation of bone apposition by osteocytes during the process of osteon refilling. Anat Rec Part A 272A:520–525, 2003. © 2003 Wiley‐Liss, Inc.
Cell and organ printing 2: Fusion of cell aggregates in three‐dimensional gels
Authors: Boland Thomas , Mironov Vladimir , Gutowska Anna , Roth Elisabeth. A. , Markwald Roger R. 
Published: May 2003

AbstractWe recently developed a cell printer (Wilson and Boland, 2003) that enables us to place cells in positions that mimic their respective positions in organs. However, this technology was limited to the printing of two‐dimensional (2D) tissue constructs. Here we describe the use of thermosensitive gels to generate sequential layers for cell printing. The ability to drop cells on previously printed successive layers provides a real opportunity for the realization of three‐dimensional (3D) organ printing. Organ printing will allow us to print complex 3D organs with computer‐controlled, exact placing of different cell types, by a process that can be completed in several minutes. To demonstrate the feasibility of this novel technology, we showed that cell aggregates can be placed in the sequential layers of 3D gels close enough for fusion to occur. We estimated the optimum minimal thickness of the gel that can be reproducibly generated by dropping the liquid at room temperature onto a heated substrate. Then we generated cell aggregates with the corresponding (to the minimal thickness of the gel) size to ensure a direct contact between printed cell aggregates during sequential printing cycles. Finally, we demonstrated that these closely‐placed cell aggregates could fuse in two types of thermosensitive 3D gels. Taken together, these data strongly support the feasibility of the proposed novel organ‐printing technology. Anat Rec Part A 272A:497–502, 2003. © 2003 Wiley‐Liss, Inc.
Development of the human Müllerian duct in the sexually undifferentiated stage
Authors: Hashimoto Ryozo 
Published: May 2003

AbstractAn embryological explanation for the development of the Müllerian duct still poses a major challenge. The development of this duct was investigated systematically in human embryos. Seven embryos (Carnegie stages 18–23) were serially sectioned in the frontal, sagittal, and transversal planes at a thickness of 10 μm and stained with hematoxylin and eosin (H&E) for histological analysis. In all observed embryos, the caudal end of the Müllerian duct was found to be intimately connected to the Wolffian duct. The opening of the Müllerian duct to the coelomic cavity was formed as the result of an invagination of the coelomic epithelium at Carnegie stage 18. The duct grew independently from the invagination during stages 19–23. The fused duct (uterovaginal canal) bifurcated at the caudal portion at Carnegie stages 22 and 23. This is the first description of the caudal portion of the fused Müllerian ducts separating again and returning to each of the Wolffian ducts in human embryos. Anat Rec Part A 272A:514–519, 2003. © 2003 Wiley‐Liss, Inc.
General suitability of techniques for in situ detection of apoptosis in small intestinal epithelium
Authors: Groos Stephanie , Reale Enrico , Luciano Liliana 
Published: May 2003

AbstractThe present study was designed to evaluate different techniques for the in situ detection of apoptosis in human and rat small intestinal epithelium. The techniques included light microscopy (LM) and transmission electron microscopy (TEM) observation of epoxy resin‐embedded tissue, scanning electron microscopy (SEM), TUNEL assay, and antibodies directed against caspase cleavage products of caspase 3, cytokeratin 18 (CK 18), and apoptotic single‐strand DNA (ssDNA). All techniques, if the labeling was positive, showed apoptotic cells exclusively at the villus tip. LM and TEM were the most reliable and revealed morphological signs typical of cells that have died via apoptosis. SEM indicated the extension of the process. The antibody recognizing cleaved caspase 3 could be considered an appropriate marker for apoptotic epithelial cells in human and rat small intestine. However, the majority of epithelial cells lining the proximal small intestinal villus contained only low levels of intact CK 18. Therefore, sufficient amounts of cleaved CK 18 for immunohistochemical detection were not generated during apoptosis, rendering the application of the antibody inappropriate. The antibody detecting formamide‐denatured ssDNA in apoptotic cells was both suitable and reliable; however, the particular staining procedure used compromised the tissue preservation. In comparison to this, the TUNEL assay was less reliable. Although it was performed with a commercially available ready‐to‐use kit, its application conditions had to be adjusted for each specimen on the basis of the findings produced by other techniques. Anat Rec Part A 272A:503–513, 2003. © 2003 Wiley‐Liss, Inc.
Morphologies and projections of defined classes of neurons in the submucosa of the guinea‐pig small intestine
Authors: Furness John B. , Alex George , Clark Melanie J. , Lal Varsha V. 
Published: May 2003

AbstractFour types of neurons have previously been identified by neurochemical markers in the submucosal ganglia of the guinea‐pig small intestine, and functional roles have been ascribed to each type. However, morphological differences among the classes have not been determined, and there is only partial information about their projections within the submucosa. In the present work, we used intracellular microelectrodes to fill neurons of each type with biocytin, which was then converted to a permanent dye, so that the shapes of the neurons could be determined and their projections within the submucosa could be followed. Cell bodies of noncholinergic secretomotor/ vasodilator neurons had Dogiel type I morphology. These neurons, which are vasoactive intestinal peptide immunoreactive, had single axons that ran through many ganglia without providing terminals around other neurons. Cholinergic secretomotor neurons with neuropeptide Y immunoreactivity had Stach type IV morphology, and cholinergic secretomotor/vasodilator neurons had stellate cell bodies. The axons of these two types ran short distances in the plexus and did not innervate other submucosal neurons. Neurons of the fourth type, intrinsic primary afferent neurons, had cell bodies with Dogiel type II morphology and their processes supplied networks of varicose processes around other nerve cells. It is concluded that each functionally defined type of submucosal neuron has a characteristic morphology and that intrinsic primary afferent neurons synapse with secretomotor neurons to form monosynaptic secretomotor reflex circuits. Anat Rec Part A 272A:475–483, 2003. © 2003 Wiley‐Liss, Inc.
Rat anterior pituitary cells in vitro can partly reconstruct in vivo topographic affinities
Authors: Noda Takahiro , Kikuchi Motoshi , Kaidzu Sachiko , Yashiro Takashi 
Published: May 2003

AbstractHormone‐producing cells in the rat anterior pituitary gland are not randomly distributed; rather, there are specific topographic affinities among five cell types (Noda et al., Acta Histochem. Cytochem. 2001;34:313–319). In this study we reconstructed these affinities, at least partially, in primary monolayer culture. Pituitary cells collected from adult male rats were enzymatically dispersed and cultured for 72 hr at a density of 1 × 105 cells/cm2. We double‐immunostained cells using antibodies against hormones, and then used confocal laser microscopy to examine the ability of the cells to attach to each other. We also statistically analyzed the affinity of all combinations of the five types of hormone‐producing cells. We observed clusters by electron microscopy to identify junctional complexes between the cells. Confocal laser microscopy indicated that the features and attachment patterns of hormone‐producing cells in vivo were similar to those in vitro. Statistical analyses revealed that the rates at which the five types of hormone‐producing cells attached to growth hormone (GH)‐, prolactin (PRL), and luteinizing hormone (LH)‐producing cells were unequal, which suggests there are specific topographic affinities. The specific rates of adrenocorticotropic hormone (ACTH)‐producing cell attachment to GH cells, LH to PRL cells, and PRL to LH cells were high, whereas that of PRL attachment to PRL cells was low. In addition, the rates correlated with the data from our previous in vivo study. Ultrastructural observations revealed few junctional complexes between hormone‐producing cells. These results indicate that anterior pituitary hormone‐producing cells can attach to specific types of cells by means of specific and/or nonspecific adhesion factors, and can reconstruct the topographic nature of the pituitary gland. Anat Rec Part A 272A:548–555, 2003. © 2003 Wiley‐Liss, Inc.
Giant mitochondria in the retina cone inner segments of shrews of genus Sorex (Insectivora, Soricidae)
Authors: Lluch Sara , López‐Fuster María José , Ventura Jacint 
Published: May 2003

AbstractThe retinas of three species of shrews (Sorex araneus, S. coronatus, and S. minutus) were analyzed. Two kinds of photoreceptors were identified according to (among other characteristics) the traits of the mitochondria of their inner segments. The rod inner segments contained several round or oval mitochondria distributed longitudinally inside the ellipsoid. The cone inner segment showed a few mitochondria, which we classified as megamitochondria (maximum length = 4.22 μm in S. araneus, 5.68 μm in S. coronatus, and 2.42 μm in S. minutus). An analysis of serial thin sections in S. coronatus showed that these large organelles occurred in the apical and central portions of the ellipsoid. In the peripheral and basal regions of the ellipsoid, megamitochondria were frequently accompanied by smaller mitochondria. The giant mitochondria were irregular in form and densely packed, and a reduced cytosol was observed between each mitochondria. In general, they exhibited an electron‐dense matrix and a complex system of cristae, which varied in length and array. In mammalian retina, megamitochondria have only been described in the ellipsoid of the tree shrews Tupaia glis and T. belangeri, two diurnal Scandentia with a rich‐cone retina. In general terms, Sorex megamitochondria are morphologically very similar to those reported for Tupaia, especially in their arrangement in the cone ellipsoid. However, they differ in the orientation of the cristae. We propose that the ellipsoid of Sorex may serve two functions: as a source of energy for receptor cells, and as a device for improving the cone outer segment optics. Anat Rec Part A 272A:484–490, 2003. © 2003 Wiley‐Liss, Inc.
Morphology of aging lung in F344/N rat: Alveolar size, connective tissue, and smooth muscle cell markers
Authors: Yamamoto Yoshio , Tanaka Akiko , Kanamaru Arata , Tanaka Shin , Tsubone Hirokazu , Atoji Yasuro , Suzuki Yoshitaka 
Published: May 2003

AbstractThis study investigated the morphological changes of lungs in F344/N rats (9–36 months old). We initially examined general and quantitative morphological changes, and then we used immunohistochemistry to detect distributional changes in collagen subtypes (types I, III, and IV) and smooth muscle cell (SMC) markers (α‐smooth muscle actin (ASMA), γ‐smooth muscle actin (GSMA), desmin, and vimentin) in the lungs. In 24‐month‐old rats, alveolar ducts and alveolar sacs were enlarged, and alveoli were wider and shallower than in younger animals. In old rats (≥27 months), terminal and respiratory bronchioles and alveolar ducts were dilated and alveoli were more extended than in 24‐month‐old rats. No age‐related distributional changes were observed for collagen types I, III, and IV as revealed by immunohistochemistry, or elastin as revealed by resorsin fuchsin. SMCs in the extra‐ and intrapulmonary bronchi were immunoreactive for ASMA, GSMA, and desmin, but not for vimentin at all ages. In old rats (≥27 months), SMCs were loosely arranged in comparison with younger animals, and stainability for GSMA and desmin was decreased. In the respiratory bronchioles and alveolar ducts, a few cells immunoreactive for ASMA and vimentin were observed in the smooth muscle aggregations of the alveolar orifice in rats younger than 12 months. In older rats (>20 months), cells immunoreactive for ASMA and vimentin were increased in septal tips. In conclusion, extension of distal airways and immunohistochemical changes of SMC markers in F344/N rat lungs were evident by ∼24 months of age, but there was no apparent change in connective tissue morphology. Anat Rec Part A 272A:538–547, 2003. © 2003 Wiley‐Liss, Inc.
Proteoglycan degradation after injurious compression of bovine and human articular cartilage in vitro: Interaction with exogenous cytokines
Authors: Patwari Parth , Cook Michael N. , DiMicco Michael A. , Blake Simon M. , James Ian E. , Kumar Sanjay , Cole Ada A. , Lark Michael W. , Grodzinsky Alan J. 
Published: May 2003

AbstractObjectiveTraumatic joint injury leads to an increased risk of osteoarthritis (OA), but the progression to OA is not well understood. We undertook this study to measure aspects of proteoglycan (PG) degradation after in vitro injurious mechanical compression, including up‐regulation of enzymatic degradative expression and cytokine‐stimulated degradation.MethodsArticular cartilage tissue explants were obtained from newborn bovine femoropatellar groove and from adult normal human donor knee and ankle tissue. Following injurious compression of the cartilage, matrix metalloproteinase 3 (MMP‐3) and MMP‐13 messenger RNA (mRNA) expression levels were measured by Northern analysis, and PG loss to the medium after cartilage injury was measured in the presence and absence of added exogenous cytokine (interleukin‐1α [IL‐1α] or tumor necrosis factor α [TNFα]).ResultsDuring the first 24 hours after injury in bovine cartilage, MMP‐3 mRNA levels increased 10‐fold over the levels in control cartilage (n = 3 experiments), whereas MMP‐13 mRNA levels were unchanged. PG loss was significantly increased after injury, but only by 2% of the total PG content and only for the first 3 days following injury. However, compared with injury alone or cytokine treatment alone, treatment of injured tissue with either 1 ng/ml IL‐1α or 100 ng/ml TNFα caused marked increases in PG loss (35% and 54%, respectively, of the total cartilage PG content). These interactions between cytokine treatment and injury were statistically significant. In human knee cartilage, the interaction was also significant for both IL‐1α and TNFα, although the magnitude of increase in PG loss was lower than that in bovine cartilage. In contrast, in human ankle cartilage, there was no significant interaction between injury and IL‐1α.ConclusionThe cytokines IL‐1α and TNFα can cause a synergistic loss of PG from mechanically injured bovine and human cartilage. By attempting to incorporate interactions with other joint tissues that may be sources of cytokines, in vitro models of mechanical cartilage injury may explain aspects of the interactions between mechanical forces and degradative pathways which lead to OA progression.
One of two chondrocyte‐expressed isoforms of cartilage intermediate‐layer protein functions as an insulin‐like growth factor 1 antagonist
Authors: Johnson Kristen , Farley David , Hu Shou‐Ih , Terkeltaub Robert 
Published: May 2003

AbstractObjectiveAging and osteoarthritic (OA) cartilage commonly demonstrate enhanced expression of the large, transforming growth factor β (TGFβ)–inducible glycoprotein cartilage intermediate‐layer protein (CILP) as well as enhanced extracellular inorganic pyrophosphate (PPi) that promotes the deposition of calcium pyrophosphate dihydrate crystals. In normal chondrocytes, TGFβ induces elevated chondrocyte extracellular PPi. Insulin‐like growth factor 1 (IGF‐1) normally blocks this response and reduces extracellular PPi. However, chondrocyte resistance to IGF‐1 is observed in OA and aging. Because CILP was reported to chromatographically fractionate with PPi‐generating nucleotide pyrophosphatase phosphodiesterase (NPP) activity, it has been broadly assumed that CILP itself has NPP activity. Our objective was to directly define CILP functions and their relationship to IGF‐1 in chondrocytes.MethodsUsing primary cultures of articular chondrocytes from the knee, we defined the function of the previously described CILP (CILP‐1) and of a recently described 50.6% identical protein that we designated the CILP‐2 isoform.ResultsBoth CILP isoforms were constitutively expressed by primary cultured articular chondrocytes, but only CILP‐1 expression was detectable in cultured knee meniscal cartilage cells. Neither CILP isoform had intrinsic NPP activity. But CILP‐1 blocked the ability of IGF‐1 to decrease extracellular PPi, an activity specific for the CILP‐1 N‐terminal domain. The CILP‐1 N‐terminal domain also suppressed IGF‐1–induced (but not TGFβ‐induced) proliferation and sulfated proteoglycan synthesis, and it inhibited ligand‐induced IGF‐1 receptor autophosphorylation.ConclusionTwo CILP isoforms are differentially expressed by chondrocytes. Neither CILP isoform exhibits PPi‐generating NPP activity. But, increased expression of CILP‐1, via N‐terminal domain–mediated inhibitory effects of CILP‐1 on chondrocyte IGF‐1 responsiveness, could impair chondrocyte growth and matrix repair and indirectly promote PPi supersaturation in aging and OA cartilage.
Epstein‐Barr virus load in the peripheral blood of patients with rheumatoid arthritis: Accurate quantification using real‐time polymerase chain reaction
Authors: Balandraud Nathalie , Meynard Jean Baptiste , Auger Isabelle , Sovran Helene , Mugnier Benedicte , Reviron Denis , Roudier Jean , Roudier Chantal 
Published: May 2003

AbstractObjectiveTo determine whether patients with rheumatoid arthritis (RA) have elevated Epstein‐Barr virus (EBV) load in their peripheral blood mononuclear cells (PBMCs) and whether it is correlated with the HLA–DR genes they express, we developed an accurate EBV DNA quantitative assay using real‐time polymerase chain reaction (PCR) with fluorescent probes.MethodsWe studied the EBV DNA load in the PBMCs of 84 patients with RA, 69 normal controls, and 22 patients with rheumatic conditions other than RA. A 214‐bp segment from the long internal repeat of EBV was amplified from 500 ng of PBMC DNA (150,000 cells) and quantified by real‐time PCR with fluorescent probes.ResultsWe demonstrated that in patients with RA, the EBV DNA load in PBMCs is increased almost 10‐fold compared with that in normal controls. The EBV load is stable over time and is not obviously influenced by disease‐modifying antirheumatic drugs or HLA–DR.ConclusionPatients with RA have elevated EBV load in their peripheral blood.
Meta‐analysis of four rheumatoid arthritis genome‐wide linkage studies: Confirmation of a susceptibility locus on chromosome 16
Authors: Fisher S. A. , Lanchbury J. S. , Lewis C. M. 
Published: May 2003

AbstractObjectiveSusceptibility to rheumatoid arthritis (RA) is likely to involve several genes of weak effect, and consequently, individual studies may have insufficient power to detect linkage. Four major RA genome‐wide linkage studies have been carried out, but apart from the well‐established HLA susceptibility locus, none of the reported significant regions of linkage has been replicated. We applied a genome‐search meta‐analysis to 4 RA genome searches to assess linkage across studies, using published results.MethodsFor each study, 120 genomic bins of ∼30 cM were defined and ranked according to the maximum evidence for linkage within each bin. Ranks were summed across studies and each bin was assessed empirically by the magnitude of summed rank, using a permutation test. A high summed rank indicated a region in which evidence for linkage was consistent across several studies.ResultsIn addition to the HLA locus (P < 0.00002), the strongest evidence for an RA susceptibility locus was found on chromosome 16 (P = 0.004). This locus was not identified as statistically significant in any of the 4 individual RA genome searches. In total, 12 regions achieved a significant (P < 0.05) summed rank, compared with the 6 bins expected by random chance. Four of these regions (on chromosomes 6p, 16cen, 6q, and 12p) reached a significance value of P < 0.01, suggesting that a subset of these regions contains RA susceptibility loci.ConclusionUsing a meta‐analysis approach, we have identified existing and novel putative RA susceptibility loci. These results can provide a basis for further positional and functional candidate‐gene studies, and may prove useful in other complex rheumatic diseases.