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Morphological Abnormalities (morphological + abnormality)
Selected AbstractsDevelopmental biology of medaka fish (Oryzias latipes) exposed to alkalinity stressJOURNAL OF APPLIED ICHTHYOLOGY, Issue 3 2010Z. L. Yao Summary Alkalinity stress is common in cultured aquatic animals and considered to be one of the major stress factors for fishes when they are transferred to saline-alkali waters. To evaluate potential effects of alkalinity on the developmental biology of Oryzias latipes, fertilized eggs, larvae and breeding fish were exposed to different carbonate alkalinity concentrations of 1.5,64.5 meq l,1, for 9, 120, and 60 days, respectively. The mortality of embryos significantly increased when exposed to the high concentrations (16.5,64.5 meq l,1). Although more than 50% of survived embryos hatched in 16.5 and 31.4 meq l,1 concentrations of carbonate alkalinity, most were not able to swim up after hatching. Morphological abnormalities such as coagulated embryos, halted embryo development, and hatching failure were observed at stages 15, 29,33 and 38 in high concentrations (31.4, 64.5 meq l,1). Almost all larvae in 16.5 and 31.4 meq l,1 treatments died 70 d post-hatch. Growth of juveniles exposed to carbonate alkalinity of 5.3 and 8.8 meq l,1 was not significantly different at 70 d and 120 d post-hatch. The number of eggs released by breeders, the fertilization rate and the hatching rate of eggs were significantly lower in the 31.4 meq l,1 treatment than in other treatments. Although medaka are capable of surviving in high alkalinities (31.4, 64.5 meq l,1) for an extended period of time, these conditions are stressful to the fish, especially at the embryonic and reproductive stages. [source] All CVB serotypes and clinical isolates induce irreversible cytopathic effects in primary cardiomyocytesJOURNAL OF MEDICAL VIROLOGY, Issue 2 2005Jeonghyun Ahn Abstract Coxsackievirus B3 (CVB3) has been identified as a major causative agent of acute and chronic myocarditis, but the involvement of other CVB serotypes in myocarditis has not been investigated. To dissect the pathological properties of different CVB serotypes toward primary cardiomyocytes, we tested their effects on primary cardiomyocyte cultures from neonatal rats. Morphological abnormalities were examined by both light and fluorescence microscopy after Hoechst 33342 staining, and loss of cell viability was estimated by MTT assay. All six CVB serotypes showed a similar degree of severe toxicity toward primary cardiomyocytes. CVB clinical isolates had cytopathic effects (CPEs) similar to those of their respective CVB reference strains. Within 1,2 days of infection with multiplicities of infection MOI 50, the cells began to experience morphological changes including cell shrinkage, rounding-up, and slight nuclear condensation. The irreversible loss of cell viability was readily observed within 3,5 days following virus infection. These results suggest that all six CVB serotypes induce direct, irreversible toxicity towards cardiomyocytes, which eventually leads to the death of infected cells. These findings indicate that the variations in CVB serotype are not the limiting factor determining the susceptibility of cardiomyocytes to CVB infection. J. Med. Virol. 75:290,294, 2005. © 2004 Wiley-Liss, Inc. [source] Ataxic mutant mice with defects in Ca2+ channel ,1A subunit gene: morphological and functional abnormalities in cerebellar cortical neuronsCONGENITAL ANOMALIES, Issue 2 2000Kazuhiko Sawada ABSTRACT This review summarizes recent studies in the morphological and functional abnormalities of cerebella in three ataxic mutant mice, i.e. tottering mouse, leaner mouse, and rolling mouse Nagoya (RMN). These mutants carry mutations in the Ca2+ channel ,1A subunit gene, and become useful models for human neurological diseases such as episodic ataxia type-2, familial hemiplegic migraine, and spinocerebellar ataxia type-6. All three mutants exhibited altered morphology of the Purkinje cells, ectopic synaptic contacts between granule cell axons (parallel fibers) and Purkinje cell dendritic spines and abnormal expression of tyrosine hydroxylase in Purkinje cells. In leaner mice, Purkinje cell loss was observed in alternating sagittal compartments of the cerebellar cortex corresponding to the Zebrin II-negative zones. The mutated Ca2+ channel ,1A subunit was highly expressed in granule and Purkinje cells, and the P-type Ca2+ currents in Purkinje cells were selectively reduced in the mutant mice. Therefore, we concluded that altered Ca2+ currents through the mutated Ca2+ channel ,1A subunit might be involved in the functional and morphological abnormalities in granule and Purkinje cells, and might result in expressions of behavioral phenotypes including ataxia. Increased levels of corticotropin-releasing factor and cholecystokinin in some climbing and mossy fibers were observed in RMN. These neuropeptides modulated the excitability of granule and Purkinje cells, indicating the possible expression of ataxic symptoms. [source] Ploidy mosaicism in well-developed nuclear transplants produced by transfer of adult somatic cell nuclei to nonenucleated eggs of medaka (Oryzias latipes)DEVELOPMENT GROWTH & DIFFERENTIATION, Issue 9 2007Elena Kaftanovskaya Chromosomal abnormalities such as ploidy mosaicism have constituted a major obstacle to the successful nuclear transfer of adult somatic cell nuclei in lower vertebrates to date. Euploid mosaicism has been reported previously in well-developed amphibian transplants. Here, we investigated ploidy mosaicisms in well-developed transplants of adult somatic cell nuclei in medaka fish (Oryzias latipes). Donor nuclei from primary cultured cells from the adult caudal fin of a transgenic strain carrying the green fluorescent protein gene (GFP) were transferred to recipient nonenucleated eggs of a wild-type strain to produce 662 transplants. While some of the transplants developed beyond the body formation stage and several hatched, all exhibited varying degrees of abnormal morphology, limited growth and subsequent death. Twenty-one transplants, 19 embryos and two larvae, were selected for chromosomal analysis; all were well-developed 6-day-old or later embryonic stages exhibiting slight morphological abnormalities and the same pattern of GFP expression as that of the donor strain. In addition, all exhibited various levels of euploid mosaicism with haploid-diploid, haploid-triploid or haploid-diploid-triploid chromosome sets. No visible chromosomal abnormalities were observed. Thus, euploid mosaicism similar to that observed in amphibians was confirmed in well-developed nuclear transplants of fish. [source] Nuclear receptor NR5A2 is required for proper primitive streak morphogenesisDEVELOPMENTAL DYNAMICS, Issue 12 2006Cassandre Labelle-Dumais Abstract NR5A2, also known as liver receptor homologue 1 (LRH-1) and fetoprotein transcription factor (FTF), is an orphan nuclear receptor involved in the regulation of cholesterol metabolism and steroidogenesis in the adult. NR5A2 was also shown to be expressed during early mouse embryogenesis. Consistent with its early expression pattern, a targeted disruption of this gene leads to embryonic lethality around the gastrulation period. To characterize the embryonic phenotype resulting from NR5A2 loss of function, we undertook morphological and marker gene analyses and showed that NR5A2,/, embryos display growth retardation, epiblast disorganization, a mild embryonic,extraembryonic constriction, as well as abnormal thickening of the proximo-posterior epiblast. We demonstrated that, although initial specification of the anterior,posterior axis occurred in the absence of NR5A2, primitive streak formation was impaired and neither embryonic nor extraembryonic mesoderm was generated. Moreover, although the visceral endoderm does not show major morphological abnormalities in NR5A2,/, embryos, a decrease in the expression level of HNF4 and GATA4 was observed. Aggregation experiments demonstrated that, in the presence of wild-type tetraploid cells, NR5A2 mutant cells in the epiblast are capable of undergoing normal gastrulation. Therefore, our results suggest a requirement for NR5A2 in extraembryonic tissues and identify a novel role of this gene in proper primitive streak morphogenesis. Developmental Dynamics 235:3359,3369, 2006. © 2006 Wiley-Liss, Inc. [source] Xenopus aristaless-related homeobox (xARX) gene product functions as both a transcriptional activator and repressor in forebrain developmentDEVELOPMENTAL DYNAMICS, Issue 2 2005Daniel W. Seufert Abstract Mutations in the aristaless-related homeobox (ARX) gene have been found in patients with a variety of X-linked mental retardation syndromes with forebrain abnormalities, including lissencephaly. Arx is expressed in the developing mouse, Xenopus, and zebrafish forebrain. We have used whole-mount in situ hybridization, overexpression, and loss-of-function studies to investigate the involvement of xArx in Xenopus brain development. We verified that xArx is expressed in the prospective diencephalon, as the forebrain is patterned and specified during neural plate stages. Expression spreads into the ventral and medial telencephalon as development proceeds through neural tube and tadpole stages. Overexpression of xArx resulted in morphological abnormalities in forebrain development, including loss of rostral midline structures, syn- or anophthalmia, dorsal displacement of the nasal organ, and ventral neural tube hyperplasia. Additionally, there is a delay in expression of many molecular markers of brain and retinal development. However, expression of some markers, dlx5 and wnt8b, was enhanced in xArx -injected embryos. Loss-of-function experiments indicated that xArx was necessary for normal forebrain development. Expansion of wnt8b expression depended on xArx function as a transcriptional repressor, whereas ectopic expression of dlx5, accompanied by development of ectopic otic structures, depended on function of Arx as a transcriptional activator. These results suggest that Arx acts as a bifunctional transcriptional regulator in brain development. Developmental Dynamics 232:313,324, 2005. © 2004 Wiley-Liss, Inc. [source] Effect of copper on hatching and development of larvae of the estuarine crab Chasmagnathus granulata (Decapoda, Brachyura)ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2001Valeria Zapata Abstract Ovigerous females of the estuarine crab Chasmagnathus granulata were exposed to 0.05, 0.1, 0.5, and 3 mg/L of copper during the egg incubation period. Regarding egg loss, a higher effect was observed at higher copper concentrations: 80% of the females lost their eggs at 0.5 mg/L, whereas no hatching was seen in those exposed to 3 mg/L. A significant decrease was found in the number of hatched larvae in females exposed to 0.5 mg/L, as was a significant decrease in the duration of the incubation period. In addition, several morphological abnormalities were seen and observed. Hydropsy and atrophy of the dorsal spine, pleon, and maxillipeds occurred at the higher copper concentrations, as found in previous studies with other pollutants. Hyperpigmentation of the cephalothorax and pleon was the only abnormality observed at every concentration assayed. Hypopigmented eyes were also evident. This pathology showed that among all defects observed at 0.5 mg/L, hypopigmented eyes had the highest incidence and might be a specific response to copper. [source] Normal embryonic development and cardiac morphogenesis in mice with Wnt1-Cre-mediated deletion of connexin43GENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 6 2006M. Kretz Abstract Mice harboring a null mutation in the gap junction protein connexin43 (Cx43) die shortly after birth due to an obstruction of the right ventricular outflow tract of the heart. These hearts exhibit prominent pouches at the base of the pulmonary outlet, i.e., morphological abnormalities that were ascribed to Cx43-deficiency in neural crest cells. In order to examine the Cx43 expression pattern in neural crest cells and derived tissues and to test whether neural crest-specific deletion of Cx43 leads to the conotruncal defects seen in Cx43null mice, we ablated Cx43 using a Wnt1-Cre transgene. Deletion of Cx43 was complete and occurred in neural crest cells as well as in neural crest-derived tissues. Nevertheless, hearts of mice lacking Cx43 specifically in neural crest cells were indistinguishable from controls. Thus, the morphological heart abnormalities of Cx43 null mice are most likely not caused by lack of Cx43 in neural crest cells. genesis 44:269,276, 2006. © 2006 Wiley-Liss, Inc. [source] The diagnosis of dysplasia and malignancy in Barrett's oesophagusHISTOPATHOLOGY, Issue 2 2000REVIEW Barrett's metaplasia is associated with an increased risk for adenocarcinoma. Adenocarcinoma develops through a multistep process characterized by defects in genes and morphological abnormalities. The early morphological changes of the process are called ,dysplasia'. Dysplasia is defined as an unequivocal neoplastic (premalignant) transformation confined within the basement membrane. For most Western pathologists malignancy is defined as invasion and characterized by a breach through the basement membrane. Japanese pathologists rely on cytological atypia and complex branching of crypts. Cytological and architectural abnormalities allow identification of dysplasia on routinely stained sections. A distinction is made between low- and high-grade dysplasia. The differential diagnosis between low-grade dysplasia and reactive changes can be difficult. Therefore a second opinion is strongly recommended, not only for high-grade dysplasia but also for low-grade. Immunohistochemistry for p53 and flow cytometry for detection of aneuploidy can support the diagnosis. Identification of dysplasia and malignancy depends on the number of biopsy samples examined. The minimum number of biopsies required has not yet been determined and depends partly on the length of the metaplastic segment. It has been proposed to sample with four quadrant biopsies at 20-mm intervals. New endoscopic techniques can increase the diagnostic yield. Endoscopically visible lesions increase the risk of finding malignancy. The time sequence for the progression of dysplasia is not known but progression from low- to high-grade and cancer has been shown to occur over a period of years although it may not be inevitable. [source] Congenital dyserythropoietic anemia type II (CDAII) is caused by mutations in the SEC23B gene,HUMAN MUTATION, Issue 9 2009Paola Bianchi Abstract Congenital dyserythropoietic anemia type II (CDAII) is an autosomal recessive disease characterized by ineffective erythropoiesis, hemolysis, erythroblast morphological abnormalities, and hypoglycosylation of some red blood cell (RBC) membrane proteins. Recent studies indicated that CDAII is caused by a defect disturbing Golgi processing in erythroblasts. A linkage analysis located a candidate region on chromosome 20, termed the CDAN2 locus, in the majority of CDAII patients but the aberrant gene has not so far been elucidated. We used a proteomic-genomic approach to identify SEC23B as the candidate gene for CDAII by matching the recently published data on the cytoplasmic proteome of human RBCs with the chromosomic localization of CDAN2 locus. Sequencing analysis of SEC23B gene in 13 CDAII patients from 10 families revealed 12 different mutations: six missense (c.40C>T, c.325G>A, c.1043A>C, c.1489C>T, c.1808C>T, and c.2101C>T), two frameshift (c.428_428delAinsCG and c.1821delT), one splicing (c.689+1G>A), and three nonsense (c.568C>T, c.649C>T, and c.1660C>T). Mutations c.40C>T and c.325G>A were detected in unrelated patients. SEC23B is a member of the Sec23/Sec24 family, a component of the COPII coat protein complex involved in protein transport through membrane vesicles. Abnormalities in this gene are likely to disturb endoplasmic reticulum (ER)-to-Golgi trafficking, affecting different glycosylation pathways and ultimately accounting for the cellular phenotype observed in CDAII. Hum Mutat 30:1,7, 2009. © 2009 Wiley-Liss, Inc. [source] Signalling mechanisms for Toll-like receptor-activated neutrophil exocytosis: key roles for interleukin-1-receptor-associated kinase-4 and phosphatidylinositol 3-kinase but not Toll/IL-1 receptor (TIR) domain-containing adaptor inducing IFN-, (TRIF)IMMUNOLOGY, Issue 3 2009Agnieszka A. Brzezinska Summary Lipopolysaccharide (LPS) stimulates exocytosis in neutrophils. The signalling molecules involved in the regulation of this mechanism are currently unknown. Using neutrophils from interleukin-1-receptor-associated kinase (IRAK)-4- and Toll/IL-1 receptor (TIR) domain-containing adaptor inducing IFN-, (TRIF)-deficient mice, we dissected the signalling pathways that control exocytosis. We analysed exocytosis of peroxidase-negative and azurophilic granules by following the mobilization of the ,2-integrin subunit CD11b and myeloperoxidase (MPO)-containing granules, respectively. IRAK-4-null neutrophils showed marked defects in both peroxidase-negative and azurophilic granule exocytosis in response to LPS. In contrast, the exocytic response to LPS of TRIF-deficient neutrophils was not different from that of wild-type cells. No differences were observed in the exocytosis of secretory organelles between IRAK-4-null and wild-type neutrophils when they were stimulated with the phorbol ester phorbol 12-myristate 13-acetate (PMA). Electron microscopy analysis showed that no morphological abnormalities were present in the granules of IRAK-4-deficient neutrophils, suggesting that the lack of exocytic response to LPS is not attributable to developmental abnormalities. Using pharmacological inhibitors, we found that p38 mitogen-activated protein kinase (p38MAPK) is essential for the exocytosis of all neutrophil secretory organelles in response to LPS. Interestingly, we found that phosphatidylinositol 3-kinase (PI3K) is essential for azurophilic granule exocytosis but not for the mobilization of other neutrophil granules in response to LPS. Azurophilic granule exocytosis in response to Listeria monocytogenes was dependent on PI3K but not IRAK-4 activity, suggesting that alternative signalling pathways are activated in IRAK-4-deficient neutrophils exposed to whole bacteria. Our results identified IRAK-4, p38MAPK and PI3K as important regulatory components with different roles in the signalling pathways that control Toll-like receptor ligand-triggered neutrophil exocytosis. [source] Quality control of bone marrow cytology; organization and over 7 years experience in the south-west NetherlandsINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 6 2008A. A. M. ERMENS Summary To asses the quality of bone marrow cytology of hospital laboratories in the south-west Netherlands a proficiency testing program was implemented. Two sets of bone marrow and blood smears from two patients were sent to 20 hospital laboratories using a tight time schedule biannually. Required results consisted of differential counts of 500 bone marrow cells and 100 peripheral blood cells, together with the description of morphological abnormalities and final conclusions. Twice a year the collected review data were discussed in a plenary session which was also used for continuous education. Over the past 7 years 30 bone marrow samples were evaluated. The coefficient of variations of specific cells counts was large. The amount of correct conclusions ranged from 12% to 100% (median: 61%). Participant attendance of the meetings was 90,100%. The total cost of this scheme of proficiency testing approximately amounted ,7000 per year. The presented formulae for both proficiency testing and haematopathological/cytological education is feasible and fulfilled the need of the participants. [source] SCANNING ELECTRON MICROSCOPY OBSERVATIONS OF DEFORMITIES IN SMALL PENNATE DIATOMS EXPOSED TO HIGH CADMIUM CONCENTRATIONS,JOURNAL OF PHYCOLOGY, Issue 6 2008Soizic Morin Different types of malformations are likely to affect the morphology of diatoms when exposed to particularly unstable environmental conditions, the most easily identifiable being distortion of the whole frustule. In the present study, we investigated, by means of SEM, valve abnormalities induced by high cadmium contamination (100 ,g · L,1) in small pennate diatoms. Changes in the shape of Amphora pediculus (Kütz.) Grunow and anomalous sculpturing of the cell wall of many species, such as Encyonema minutum (Hilse) D. G. Mann, Mayamaea agrestris (Hust.) Lange-Bert., Gomphonema parvulum (Kütz.) Kütz., or Eolimna minima (Grunow) Lange-Bert., were observed, which were not, or almost not, noticeable in the LM. With consideration to current knowledge of diatom morphogenesis, metal uptake by the cell would induce, directly or indirectly, damage to many cytoplasmic components (e.g., microtubules, cytoskeleton, Golgi-derived vesicles) involved in the precisely organized silica deposition. This study confirms that many species, whatever their size, are likely to exhibit morphological abnormalities under cadmium stress, and that this indicator may be valuable for the biomonitoring of metal contamination, even if SEM observations are not necessary for routine studies. [source] QUANTITATIVE SENSORY TESTING AND SWEAT FUNCTION IN FRIEDREICH'S ATAXIA.JOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2000CORRELATION WITH CUTANEOUS INNERVATION To evaluate small fiber function in Friedreich's Ataxia (FA), we performed in 7 patients pin-prick, thermal thresholds, and sweat test. All tests were performed in four different sites: hand dorsum, anterior thigh, lateral distal leg, and foot dorsum. The same subjects underwent 3 mm punch skin biopsy from fingertip, anterior thigh, and lateral distal leg. We used a thin needle mounted on a calibrated nylon wire for the pin-prick test, and a Medoc 2001 TSA system for thermal threshold assessment. Sweat test was performed using a silicon mold after stimulation with pilocarpine by iontophoresis. Skin specimens, cut into 100-,m-thick sections, were double-stained using primary antibodies specific for collagen and nervous fibers and secondary antibodies labeled with Cy3 and Cy5 fluorophores. Tridimensional digitized images were obtained from z-series of 2-,m-thick optical sections acquired with a confocal microscope. We found in all patients in the more distal sites definite signs of functional impairment of the small fibers. These data correlated with the skin innervation morphological findings that showed, in the same sites, a sensible loss of small fibers regarding both the epidermal free endings and the subepidermal neural plexus. Less severe morphological abnormalities were found in the proximal sites. The large fiber neuropathy in FA is well documented. Our data show a length-dependent involvement of small fibers in the pathological process. [source] Branching sites and morphological abnormalities behave as ectopic poles in shape-defective Escherichia coliMOLECULAR MICROBIOLOGY, Issue 4 2004Trine Nilsen Summary Certain mutants in Escherichia coli lacking multiple penicillin-binding proteins (PBPs) produce misshapen cells containing kinks, bends and branches. These deformed regions exhibit two structural characteristics of normal cell poles: the peptidoglycan is inert to dilution by new synthesis or turnover, and a similarly stable patch of outer membrane caps the sites. To test the premise that these aberrant sites represent biochemically functional but misplaced cell poles, we assessed the intracellular distribution of proteins that localize specifically to bacterial poles. Green fluorescent protein (GFP) hybrids containing polar localization sequences from the Shigella flexneri IcsA protein or from the Vibrio cholerae EpsM protein formed foci at the poles of wild-type E. coli and at the poles and morphological abnormalities in PBP mutants. In addition, secreted wild-type IcsA localized to the outer membrane overlying these aberrant domains. We conclude that the morphologically deformed sites in these mutants represent fully functional poles or pole fragments. The results suggest that prokaryotic morphology is driven, at least in part, by the controlled placement of polar material, and that one or more of the low-molecular-weight PBPs participate in this process. Such mutants may help to unravel how particular proteins are targeted to bacterial poles, thereby creating important biochemical and functional asymmetries. [source] Double-stranded RNA-mediated gene silencing of cysteine proteases (falcipain-1 and -2) of Plasmodium falciparumMOLECULAR MICROBIOLOGY, Issue 5 2002Pawan Malhotra Summary Malaria remains a public health problem of enormous magnitude, affecting over 500 million people every year. Lack of success in the past in the development of new drug/vaccines has mainly been attributed to poor understanding of the functions of different parasite proteins. Recently, RNA interference (RNAi) has emerged as a simple and incisive technique to study gene functions in a variety of organisms. In this study, we report the results of RNAi by double-stranded RNA of cysteine protease genes (falcipain -1 and -2) in the malaria parasite, Plasmodium falciparum. Using RNAi directed towards falcipain genes, we demonstrate that blocking the expression of these genes results in severe morphological abnormalities in parasites, inhibition of parasite growth in vitro and substantial accumulation of haemoglobin in the parasite. The inhibitory effects produced by falcipain double-stranded (ds)RNAs are reminiscent of the effects observed upon administering E-64, a cysteine protease inhibitor. The parasites treated with falcipain's dsRNAs also show marked reduction in the levels of corresponding endogenous falcipain mRNAs. We also demonstrate that dsRNAs of falcipains are broken into short interference RNAs , 25 nucleotides in size, a characteristic of RNAi, which in turn activates sequence-specific nuclease activity in the malaria parasites. These results thus provide more evidence for the existence of RNAi in P. falciparum and also suggest possibilities for using RNAi as an effective tool to determine the functions of the genes identified from the P. falciparum genome sequencing project. [source] Sperm defects in mice lacking a functional Niemann,Pick C1 protein,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 10 2006Jun Fan Abstract The Niemann,Pick C1 (NPC1) gene encodes for a multiple membrane spanning protein, which regulates the trafficking of low-density lipoprotein-mediated endocytosed cholesterol. Mutation of the human NPC1 gene causes Niemann,Pick type C (NPC) disease. The Npc1NIH mice, a model of human NPC disease, bear a spontaneous mutation of the Npc1 gene, and are infertile. In this study, we have performed sperm analysis to search for the cause of male infertility in the Npc1NIH mouse. The number of cauda sperms in Npc1,/, mice was decreased roughly three-and-half-fold of that in wild-type mice. The decreased sperm number in Npc1,/, mice is due, at least in part, to partial arrest of spermatogenesis in the testes, as revealed by histological analysis. Compared to wild-type sperm, Npc1,/, sperm displayed a high frequency of morphological abnormalities, including tailless heads and aberrant heads. In the in vitro fertilization (IVF) assay using cumulus-intact eggs, Npc1,/, sperm failed to produce two-cell embryos. In the IVF assay where zona-free eggs were used, Npc1,/, sperm bound normally but could not fuse with the egg. Further analysis indicated that Npc1,/, sperms are drastically impaired in the binding to the egg zona pellucida, only 14% of the level of wild-type sperm. Moreover, on Npc1,/, cauda sperm, one-third of the total cyritestin protein was not proteolytically processed, while fertilin , was processed normally. Taken together, these results demonstrate that there are multiple defects in sperms from mice lacking a functional NPC1 protein, and these observed sperm defects may result in sterility. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source] Effect of pyriproxyfen, a juvenile hormone mimic, on egg hatch, nymph development, adult emergence and reproduction of the Asian citrus psyllid, Diaphorina citri KuwayamaPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 4 2010Dhana Raj Boina Abstract BACKGROUND: The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is a vector of bacteria presumably responsible for huanglongbing (HLB) disease in citrus. In this laboratory study, an investigation was made of the activity of pyriproxyfen, a juvenile hormone mimic, on ACP eggs, nymphs and adults to evaluate its potential as a biorational insecticide for inclusion in an integrated pest management (IPM) program for ACP. RESULTS: Irrespective of egg age, timing or method of treatment, a significantly lower percentage of eggs (5,29%) hatched after exposure to 64 and 128 µg mL,1 of pyriproxyfen. Only 0,36% of early instars (first, second and third) and 25,74% of late instars (fourth and fifth) survived to adults following exposure to 16, 32 and 64 µg mL,1 of pyriproxyfen. However, 15,20% of adults that emerged following treatment as late instars exhibited morphological abnormalities. Furthermore, pyriproxyfen adversely affected reproduction (fecundity and fertility) of adults that emerged from treated fifth instars or that were treated topically with 0.04 µg as adults. CONCLUSIONS: Application of pyriproxyfen at 64 µg mL,1 resulted in greater inhibition of egg hatch and suppression of adult emergence compared with lower rates. Pyriproxyfen also markedly reduced female fecundity and egg viability for adults that were exposed either as fifth instars or as newly emerged adults. The ovicidal, larvicidal and reproductive effects against ACP suggest that pyriproxyfen is suitable for integration into an IPM program for ACP. Copyright © 2009 Society of Chemical Industry [source] Identification of differentially expressed genes related to aberrant phenotypes in Brassica oleracea var. botrytisPLANT BREEDING, Issue 6 2009A. Salmon Abstract The aberrant phenotype is characterized by the progressive expression of abnormal traits during vegetative growth affecting leaf thickness, shape and/or plant vigour. These striking morphological abnormalities do not appear to be caused by agronomical practices or pathogen infections. Furthermore, the aberrant phenotype, which is observed in 3,20% of cultivated cauliflowers, is not linked to DNA sequence or structural polymorphisms. To detect candidate genes related to the aberrant phenotype, we used amplified fragment length polymorphism on cDNA approach, sampling normal and aberrant F1 hybrid plants several times before and after the expression of the aberrant phenotype. This screen led to the detection of 51 differentially expressed transcripts. Twenty-nine of these were homologous to annotated genes in genomic databases. We identified transcripts, which were differentially expressed before the expression of the aberrant trait with homology to genes involved in various abiotic stress responses. A non-specific lipid transfer protein homologue was also identified and given the role that these proteins play in epicuticular wax formation and leaf morphology, it may be implicated in the abnormal leaf shape phenotypes. [source] Over-expression of a Populus peroxisomal ascorbate peroxidase (PpAPX) gene in tobacco plants enhances stress tolerancePLANT BREEDING, Issue 4 2009Y-J. Li Abstract Ascorbate peroxidase (APX) plays an important role in the metabolism of hydrogen peroxide in higher plants. We studied the effect of over-expressing a Populus peroxisomal ascorbate peroxidase (PpAPX) gene under the control of the cauliflower mosaic virus 35S promoter or the rd29 promoter in transgenic tobacco. High levels of PpAPX gene expression were observed in 35S-PpAPX transgenic plants, with a 50% increase in APX activity. The constitutive expression of PpAPX in the tobacco exhibited no morphological abnormalities, while significantly increased root growth was observed in transgenic plants, when compared to control plants. Several independently transformed lines were propagated and evaluated for resistance to methyl viologen (MV), drought and salt stress. Visual assessment of transgenic and control lines exposed to MV (50 or 100 ,mol) confirmed that over-expression of APX minimized leaf damage. APX activity was nearly 80% higher in the leaves of transgenic plants in response to drought or salt stresses. Moreover, the transgenic tobacco also showed significantly improved drought resistance and salt tolerance at the vegetative stage. RNA blot analysis indicated that the PpAPX transcript level was very low under normal growing conditions in rd29Ap-PpAPX plants, but clearly increased under drought stress. Our results show that PpAPX does not play a significant role under normal growing conditions, but did ameliorate oxidative injury under abiotic stress. The Ad29 promoter should be used as an inducible promoter in transgenic works. [source] Inhibitory effects of furanone metabolites of a rhizobacterium, Pseudomonas jessenii, on phytopathogenic Aphanomyces cochlioides and Pythium aphanidermatumPLANT PATHOLOGY, Issue 1 2010A. Deora An antagonistic rhizobacterium, Pseudomonas jessenii EC-S101, isolated from the rhizosphere of spinach, produces two related secondary metabolites, 3-[(1R)-hydroxyoctyl]-5-methylene-2(5H)-furanone (4,5-didehydroacaterin) (1) and 3-[(1R)-hydroxyhexyl]-5-methylene-2(5H)-furanone (2). This study demonstrated their in vitro inhibitory effects, in particular those of (1), against Aphanomyces cochlioides AC-5 and Pythium aphanidermatum PA-5. The compounds inhibited radial growth and induced morphological abnormalities characterized by hyperbranching and periodic swelling in AC-5 and PA-5 hyphae, respectively. Staining with rhodamine-phalloidin, which binds to plasma-membrane-associated filamentous-actin (F-actin), revealed that tip-specific actin filaments were remodelled into a plaque-like form at an early stage of encounter (up to 24 h) with (1) or (2), whereas at later stages of encounter (48 h), the plaques were eliminated, reflecting the disorganization of actin arrays in the morphologically abnormal AC-5 and PA-5 hyphae. A similar response of actin disorganization was observed in AC-5 and PA-5 hyphae upon treatment with latrunculin B (3), an actin-assembly inhibitor produced by a sea sponge. It is suggested that (1) and (2) caused actin disorganization and their inhibitory activities were comparable to that of (3). Further ultrastructural observations substantiated abnormal functioning and delocalization of F-actin-linked cell organelles. [source] Comparison of the Effect of the Aromatase Inhibitor, Anastrazole, to the Antioestrogen, Tamoxifen Citrate, on Canine Prostate and SemenREPRODUCTION IN DOMESTIC ANIMALS, Issue 2009G Gonzalez Contents This study compared the efficiency of the aromatase inhibitor, anastrazole, with the antioestrogenic receptor blocker, tamoxifen, on normal (NRL) and hyperplastic prostate glands. Forty healthy dogs were classified as NRL (n = 18) or abnormal (ABN) with benign prostate hyperplasia (n = 22). The dogs were randomly assigned to one of the following six groups, treated for 60 days; oral placebo for normal (NRL-PLC; n = 6) and abnormal (ABN-PLC; n = 6), oral anastrazole 0.25,1 mg/day, for normal (NRL-ANZ, n = 6) and abnormal (ABN-ANZ, n = 8) and oral tamoxifen citrate 2.5,10 mg/day for normal (NRL-TMX; n = 6) and abnormal (ABN-TMX; n = 8) dogs. The dogs were evaluated before treatment and then monthly for 4 months. At the end of the treatment, the prostatic volume decreased by 28.5 ± 4.3%, 21.6 ± 6.3% and 0.7 ± 1.0% in the ABN-TMX, ABN-ANZ and ABN-PLC (p < 0.01), respectively. From then on, prostatic volume began to increase without reaching pre-treatment values at the end of the study. In the ABN animals, there were no differences for this parameter between ANZ and TMX treatment (p > 0.1), whereas in the NRL animals ANZ produced a less pronounced decrease (p < 0.05), libido, testicular consistency and scrotal diameter decreased during treatment in the TMX group (p > 0.05). These parameters and sperm volume, count, motility and morphological abnormalities remained unaltered throughout the study in the ANZ and PLC groups (p > 0.05). There were no haematological nor biochemical side effects. Anastrazole might offer a safe and effective alternative for the medical management of dogs with benign prostatic hyperplasia. [source] Obstructive infertility: changes in the histology of different regions of the epididymis and morphology of spermatozoaANDROLOGIA, Issue 4 2006P. C. Pal Summary In the present study, the effects of prolonged obstruction in different regions of the human epididymis on its histology and on the spermatozoa retained at the site of obstruction were assessed. Men who were confirmed of having obstruction of the epididymis underwent vasoepididymostomy (VEA) for surgical correction of the obstruction. At the time of surgery, fluid from the epididymal tubule above the site of obstruction was aspirated and examined for sperm profile. Epididymal tissue, collected at the site of obstruction, was processed for assessment of histological changes and also used to identify the site of obstruction. Prolonged obstruction of the epididymis has caused degeneration of the epididymal epithelium, gradual decrease in the diameter of the tubule and tubular lumen and increase in the intertubular connective tissue. Sperm aspirated from the caput epididymal fluid showed sluggish pattern of motility only in one out of the six subjects, whereas spermatozoa collected from the cauda epididymal fluid showed rapid linear progressive motility in one of three subjects. A major percentage of spermatozoa in the aspirated fluid showed various types of morphological abnormalities, irrespective of the site of obstruction. These results are discussed in relation to the role of the epididymis in investing spermatozoa with motility and fertilizing capacity. [source] Perinatal outcome in fetuses with extremely large nuchal translucency measurementAUSTRALIAN AND NEW ZEALAND JOURNAL OF OBSTETRICS AND GYNAECOLOGY, Issue 3 2009Fergus SCOTT Background: Studies have suggested that an entirely normal outcome is likely when the nuchal translucency (NT) measurement is very large and the karyotype, morphology and echocardiography scans are normal. Recently this has been questioned as it is based on very small numbers. Aim: Assess the outcome of pregnancies with an NT measurement of 6.5 mm or greater. Methods: Audit of a large first trimester screening program. Results: Over the ten years to 2006, 76 813 patients underwent first trimester screening, with 120 having an extremely large NT. Thirty-one cases had normal karyotypes, of which there were four sets of twins that demised. Six cases miscarried and ten were terminated, some with morphological abnormalities. Eight cases were still alive for the morphology scan, with the only abnormality being mild pyelectasis in one case. At birth, three cases were normal and another three cases had a good outcome. Two cases had coarctation of the aorta and a good outcome. One case had Noonan's syndrome, another had cerebral palsy and the case with pyelectasis had hydronephrosis, dilated ureters and some contractures. Conclusions: When the karyotype and morphology scan are normal, the outcome is often good in spite of an extremely large NT. However, even a subtle ultrasound anomaly can indicate a genetic syndrome and echocardiography cannot exclude mild cardiac abnormalities. [source] Sulphated Polysaccharides: New Insight in the Prevention of Cyclosporine A-Induced Glomerular InjuryBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 1 2007Anthony Josephine Nephrotoxicity induced by cyclosporine A continues to be a major problem despite its potent immunosuppressive action. Adult male albino rats of Wistar strain were categorized into four groups. Two groups (II and IV) were administered cyclosporine A (25 mg/kg body weight, orally) for 21 days, in which Group IV rats were also treated simultaneously with sulphated polysaccharides (5 mg/kg body weight, subcutaneously) for the same period. A significant loss in body weight was noted in the cyclosporine A-induced rats. Renal damage was assessed in terms of decreased creatinine clearance and increased activity of lysosomal enzymes. The levels of glycoproteins were found to be decreased in the renal tissue, and a noticeable rise in glycosaminoglycanuria coupled with marked proteinuria was more prominent in the cyclosporine A-induced animals. Furthermore, the extent of kidney damage was assessed by histopathological findings. Toxic manifestations were also confirmed by transmission electron microscopic studies. These morphological abnormalities and other alterations in the renal tissue were significantly offset by sulphated polysaccharides supplementation. These findings underline that restoration of normal cells accredits sulphated polysaccharides, from Sargassum wightii, with nephroprotective role, against cyclosporine A-induced renal injury. [source] PERCUTANEOUS TRANSCATHETER CLOSURE OF PATENT DUCTUS ARTERIOSUS WITH AN AMPLATZER DUCT OCCLUDER USING RETROGRADE GUIDEWIRE-ESTABLISHED FEMORAL ARTERIOVENOUS LOOPCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 5-6 2008Jian-Fa Zhang SUMMARY 1The traditional antegrade wire-guided percutaneous transcatheter approach is not ideal in closing some types of patent ductus arteriosus (PDA) with abnormal morphology. The aim of the present study was to evaluate the efficacy of a retrograde wire-guided transcatheter approach for closure of some types of PDA using the Amplatzer duct occluder (ADO). 2Nineteen patients with abnormal PDA morphology, including a smaller ostium of the side of the pulmonary artery compared with the side of the descending aorta, severe calcification or tortuosity, were included in the present study. In these patients, after the antegrade approach failed to cross a wire from the pulmonary artery via the PDA to the descending aorta, a retrograde guidewire was passed through the PDA in the opposite direction, from the descending aorta to the pulmonary artery, to establish a femoral arteriovenous loop that assisted the deployment of the ADO in all 19 patients. The size of the PDA, as determined by angiography, was 3.1 ± 1.1 mm and the diameter of the ADO selected was 6.5 ± 1.5 mm. 3In 16 cases, systolic murmur disappeared after the procedure. Systolic murmur (less than Grade II) and angiographic residual shunt remained in three cases immediately after the procedure, but disappeared 1 month later. Mean pulmonary arterial pressure decreased from 33 ± 8 to 22 ± 4 mmHg in all 19 patients (P < 0.01). There were no complications during or after the procedure. 4The retrograde wire-guided technique offers an alternative approach to facilitate closure of a PDA that cannot be achieved by traditional antegrade wire-guided methods due to morphological abnormalities in the PDA. [source] Alcoholic macrocytosis,is there a role for acetaldehyde and adducts?ADDICTION BIOLOGY, Issue 1 2004Onni Niemelä Although alcohol abuse is known to cause a wide array of adverse effects on blood cell formation, the molecular mechanisms by which alcohol exerts its toxic actions have remained poorly defined. Elevated mean corpuscular volume (MCV), macrocytosis, is the most typical morphological abnormality induced by excessive ethanol consumption. This paper reviews recent data indicating that acetaldehyde, the first metabolite of ethanol, may play a role in the haematological derangements in peripheral blood cells and in bone marrow of alcoholic patients. Studies in experimental animals and in human alcoholics have shown that acetaldehyde can bind to proteins and cellular constituents forming stable adducts. Elevated adduct levels have been found from the erythrocytes of alcohol abusers, which may also be associated with ethanol-induced effects in haematopoiesis and adverse consequences in cellular functions. [source] Characterization of teleost Mdga1 using a gene-trap approach in medaka (Oryzias latipes)GENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 8 2009Shinya Sano Abstract MAM domain containing glycosilphosphatidilinositol anchor 1 (MDGA1) is an IgCAM protein present in many vertebrate species including humans. In mammals, MDGA1 is expressed by a subset of neurons in the developing brain and thought to function in neural cell migration. We identified a fish ortholog of mdga1 by a gene-trap screen utilizing the Frog Prince transposon in medaka (Japanese killifish, Oryzias latipes). The gene-trap vector was inserted into an intronic region of mdga1 to form a chimeric protein with green fluorescent protein, allowing us to monitor mdga1 expression in vivo. Expression of medaka mdga1 was seen in various types of embryonic brain neurons, and specifically in neurons migrating toward their target sites, supporting the proposed function of MDGA1. We also isolated the closely related mdga2 gene, whose expression partially overlapped with that of mdga1. Despite the fact that the gene-trap event eliminated most of the functional domains of the Mdga1 protein, homozygous embryos developed normally without any morphological abnormality, suggesting a functional redundancy of Mdga1 with other related proteins. High sequential homology of MDGA proteins between medaka and other vertebrate species suggests an essential role of the MDGA gene family in brain development among the vertebrate phylum. genesis 47:505,513, 2009. © 2009 Wiley-Liss, Inc. [source] Intramitochondrial crystalline inclusions in nonalcoholic steatohepatitis,HEPATOLOGY, Issue 6 2009Stephen H. Caldwell Mitochondrial dysfunction is an important element in the pathogenesis of nonalcoholic steatohepatitis (NASH). Intramitochondrial crystals (IMCs) are a well-documented morphological abnormality seen on transmission electron microscopy in this disease. It has been suggested that IMCs consist of phospholipids, but their exact composition remain uncertain many years after their discovery. Micellar phase transitions of phospholipid bilayers is a well-known but little-studied phenomenon in living systems. Its presence in the mitochondria of NASH would offer significant insight into the disease with possible therapeutic implications. We postulated that intramitochondrial disturbances in NASH are sufficient to produce such transitions and that their detection in fresh biopsies would therefore be a dynamic process. To test this, we performed a blinded, prospective analysis of fresh liver biopsy samples immediately fixed under different conditions. Quantitative transmission electron microscopy morphometry, performed by systematically counting total mitochondria and IMCs within areas of uniform dimension, showed a stepwise decline in IMCs with cooler fixation temperature in each subject studied. Randomization testing (Monte Carlo resampling) confirmed that the detection of IMCs was strongly dependent on fixation temperature (P < 0.0001). Conclusion: These results indicate that the intramitochondrial crystals characteristic of NASH are highly dynamic and unstable structures. The findings offer the strongest support yet for their origin in micellar phase transitions. We speculate that such transitions result from microenvironmental changes within the mitochondria and carry therapeutic implications, especially in regard to dietary manipulations of mitochondrial lipid composition. (HEPATOLOGY 2009.) [source] Performance of the XE-2100 leucocyte differentialINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 5 2002G. Stamminger Summary The XE-2100Ō was evaluated in a multicentre study following a previously established protocol. In this paper, we demonstrate the results of analytical performance studies, including comparison of the leucocyte differential with the NCCLS H20-A method and evaluation of flagging sensitivity. Linearity of the leucocyte count over a wide clinical range, low imprecision in clinically important ranges and no measurable carry over were confirmed. For comparability studies, 4 × 200 cell microscopic differential leucocyte counts were correlated with the automated five-part-differential counts. No significant differences were detected in (1) a group without morphological abnormality and in (2) a leukopenic group. The sensitivity of flags for the detection of immature granulocytes and myeloid blasts was very good. Only few samples containing blast cells remained unrecognized but these would have been examined microscopically in any event because of other abnormalities indicated by the instrument. Atypical/abnormal lymphocytes/and lymphoblasts were detected very reliably when the total lymphocyte count and the flags were evaluated in combination. A similiar procedure is recommended for the detection of left shift. When the neutrophil count is elevated, the sensitivity of the left shift flag is improved. The absolute immature granulocyte (IG) count by the instrument correlates well with that of myeloid precursor cells by microscopy. [source] | ||||||||||||||||||||||||||||