DERMATOLOGIC SURGERY, Issue 6 2004
Robert A. Schwartz MD
Background: Cutaneous metastatic disease is an important finding that may represent the first sign of systemic cancer, or, if already known, that may change tumor staging and thus dramatically altered therapeutic plans. Although cutaneous metastases are relatively frequent in patients with cutaneous melanoma, they are less so from ocular melanoma. Objective: To demonstrate the value of HMB-45, staining in the detection of ocular melanoma metastatic to skin. Methods: The immunohistochemical stain HMB-45 a monoclonal antibody directed against intact human melanoma cells, was employed on a skin biopsy specimen from a cutaneous tumor. Results: HMB-45 staining was positive in the atypical hyperchromatic cells of the deep dermis. Conclusion: HMB-45 may be of value in the detection of ocular melanoma metastatic to skin. Cutaneous metastatic disease is a somewhat common and extremely important diagnosis. Although cutaneous metastases from cutaneous melanoma are relatively frequent, those from ocular melanomas are less so. Use of histochemical staining, especially the HMB-45 stain, allows confirmation of the diagnosis. [source]

Immunocytochemical study of the expression of mesothelin in fine-needle aspiration biopsy specimens of pancreatic adenocarcinoma

DIAGNOSTIC CYTOPATHOLOGY, Issue 3 2007
Amy C. Baruch M.D.
Abstract Mesothelin is a potential marker of pancreatic adenocarcinoma that was recently identified by serial analysis of gene expression. We evaluated the sensitivity and specificity of mesothelin as a marker of pancreatic adenocarcinoma on destained Papanicolaou (Pap) smears and unstained cellblocks from 28 patients using a monoclonal antibody to mesothelin. Intensity and proportion of staining was semiquantitatively graded on a scale of 1,3, and as 0%, 1 to <10%, 10,50%, or >50%. Positive staining for mesothelin was seen in 64% of the direct smears and in 36% of cell block sections. Focal positivity for mesothelin was noted in benign pancreatic tissue in one of 10 cases. Staining was most often focal (<50% of cells) in both direct smears and cell block sections. The overall sensitivity and specificity of mesothelin as a marker for pancreatic adenocarcinoma were 68% and 90%, respectively. Sensitivity was higher in Pap smears than in cell block sections (64% versus 36%). The presence of occasional mesothelin expression in benign tissue, its very focal expression in malignant tissue may limit the utility of mesothelin as a marker of pancreatic adenocarcinomas in fine-needle aspiration (FNA) specimens. Diagn. Cytopathol. 2007;35:143,147. © 2007 Wiley-Liss, Inc. [source]

Availability of autoantigenic epitopes controls phenotype, severity, and penetrance in TCR Tg autoimmune gastritis

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 12 2008
Ditza Levin
Abstract We examined TCR:MHC/peptide interactions and in vivo epitope availability to explore the Th1- or Th2-like phenotype of autoimmune disease in two TCR Tg mouse models of autoimmune gastritis (AIG). The TCR of strains A23 and A51 recognize distinct IAd -restricted peptides from the gastric parietal cell H/K-ATPase. Both peptides form extremely stable MHC/peptide (MHC/p) complexes. All A23 animals develop a Th1-like aggressive, inflammatory AIG early in life, while A51 mice develop indolent Th2-like AIG at 6,8,wk with incomplete penetrance. A51 T cells were more sensitive than A23 to low doses of soluble antigen and to MHC/p complexes. Staining with IAd/peptide tetramers was only detectable on previously activated T cells from A51. Thus, despite inducing a milder AIG, the A51 TCR displays a higher avidity for its cognate IAd/peptide. Nonetheless, in vivo proliferation of adoptively transferred A51 CFSE-labeled T cells in the gastric lymph node was relatively poor compared with A23 T cells. Also, DC from WT gastric lymph node, presenting processed antigen available in vivo, stimulated proliferation of A23 T cells better than A51. Thus, the autoimmune potential of these TCR in their respective Tg lines is strongly influenced by the availability of the peptide epitope, rather than by differential avidity for their respective MHC/p complexes. [source]

Short exposure to high levels of fluoride induces stage-dependent structural changes in ameloblasts and enamel mineralization

EUROPEAN JOURNAL OF ORAL SCIENCES, Issue 2006
D. M. Lyaruu
We tested the hypothesis that the sensitivity of forming dental enamel to fluoride (F,) is ameloblast developmental stage-dependent and that enamel mineralization disturbances at the surface of fluorotic enamel are caused by damage to late-secretory- and transitional-stage ameloblasts. Four-day-old hamsters received a single intraperitoneal dose of 2.5,20 mg NaF/kg body weight and were examined, 24 h later, by histology and histochemistry. A single dose of ,,5 mg of NaF/kg induced the formation of a hyper- followed by a hypomineralized band in the secretory enamel, without changing the ameloblast structure. At 10 mg of NaF/kg, cystic lesions became apparent under isolated populations of distorted late-secretory- and transitional-stage ameloblasts. Staining with von Kossa stain showed that the enamel under these lesions was hypermineralized. At 20 mg of NaF/kg, cystic lesions containing necrotic cells were also found in the early stages of secretory amelogenesis and were also accompanied with hypermineralization of the enamel surface. We concluded that the sensitivity to F, is ameloblast developmental stage-dependent. Groups of transitional ameloblasts are most sensitive, followed by those at early secretory stages. These data suggest that a F-induced increase in cell death in the transitional-stage ameloblasts accompanies the formation of cystic lesions, which may explain the formation of enamel pits seen clinically in erupted teeth. [source]

Generation and characterization of Csrp1 enhancer-driven tissue-restricted Cre-recombinase mice

GENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 5 2008
Paige Snider
Histological section showing beta-galactosidase staining (blue) of embryonic day 12.5 heart from a new cysteine-rich protein (Cspr1) transgenic mouse strain crossed to the R26R cre reporter mouse. Staining is found specifically in the outflow tract cushions and myocardial cuff. See the paper by Snider et al. in the March 2008 issue. [source]

Irreversible electroporation of the pancreas in swine: a pilot study

HPB, Issue 5 2010
Kevin P. Charpentier
Abstract Background:, Irreversible electroporation (IRE) is a novel, non-thermal method of tissue ablation using short pulses of high-voltage DC current to ablate tissue. Methods:, Irreversible electroporation of the pancreas was performed in four domestic female swine using two monopolar probes spaced 9,15 mm apart. Ninety pulses of 1500 V/cm were delivered for each ablation. Results:, All animals survived for their designated times of 2 h (n= 1), 2 days (n= 1) and 14 days (n= 2), respectively. No procedure-related complications occurred. Three animals in which probes had been spaced at intervals of 10 ± 1 mm showed evidence of irreversible ablation, with ablation height ranging from <10 mm to 21 mm and ablation width ranging from <10 mm to 16 mm by gross appearance and triphenyltetrazolium chloride (TTC) staining. The only animal in which probes had been spaced at intervals of 15 mm did not show evidence of irreversible ablation at 2 weeks. This may be secondary to the wider probe spacing and relatively low voltage, which results in a mostly reversible form of electroporation without cell death. Conclusions:, Irreversible electroporation appears to be a safe method for pancreas tissue ablation. Staining with TTC can predict the zone of IRE ablation within 2 h of treatment. [source]

MAGE-A9 mRNA and protein expression in bladder cancer

INTERNATIONAL JOURNAL OF CANCER, Issue 10 2007
Valérie Picard
Abstract In a previous analysis, we showed that MAGE-As were the most frequently expressed cancer-testis antigens in human bladder tumours. Here, we further characterized by RT-PCR the expression of this family of genes by analyzing specifically MAGE-A3, -A4, -A8 and -A9 mRNAs in 46 bladder tumours and 10 normal urothelia. We found that they were expressed in 30, 33, 56 and 54% of tumours, respectively. Although MAGE-A8 was the most frequent, its expression was low and was also found in most normal urothelia. The other MAGE-A mRNAs were all tumour-specific but MAGE-A9 mRNA was expressed at a higher level and was two times more frequent in superficial than in invasive tumours. To study the expression of the protein, we produced 2 MAGE-A9-specific monoclonal antibodies (mAbs) presenting no cross-reactivity with other MAGE-A proteins. MAb 14A11, was used to analyse the expression of the antigen in testis and tumour samples by immunohistochemistry. In testis, MAGE-A9 expression was restricted to primary spermatocytes. Most bladder tumours that expressed the MAGE-A9 transcript were positive with mAb 14A11. Staining was heterogeneous but half of the tumours showed over 75% positive cells. Finally, we showed that treatment of bladder cancer cells with the methylation inhibitor, 5-aza-2,-deoxycytidine, alone or in combination with the histone deacetylase inhibitors MS-275 and 4-phenylbutyrate could strongly induce the expression of MAGE-A9. These results show that MAGE-A9 is frequently expressed in superficial bladder cancer and could be a relevant target for immunotherapy or chemoimmunotherapy because its expression can be induced by chemotherapeutic drugs. © 2007 Wiley-Liss, Inc. [source]

A comparative study of gland cells implicated in the nerve dependence of salamander limb regeneration

JOURNAL OF ANATOMY, Issue 1 2010
Anoop Kumar
Abstract Limb regeneration in salamanders proceeds by formation of the blastema, a mound of proliferating mesenchymal cells surrounded by a wound epithelium. Regeneration by the blastema depends on the presence of regenerating nerves and in earlier work it was shown that axons upregulate the expression of newt anterior gradient (nAG) protein first in Schwann cells of the nerve sheath and second in dermal glands underlying the wound epidermis. The expression of nAG protein after plasmid electroporation was shown to rescue a denervated newt blastema and allow regeneration to the digit stage. We have examined the dermal glands by scanning and transmission electron microscopy combined with immunogold labelling of the nAG protein. It is expressed in secretory granules of ductless glands, which apparently discharge by a holocrine mechanism. No external ducts were observed in the wound epithelium of the newt and axolotl. The larval skin of the axolotl has dermal glands but these are absent under the wound epithelium. The nerve sheath was stained post-amputation in innervated but not denervated blastemas with an antibody to axolotl anterior gradient protein. This antibody reacted with axolotl Leydig cells in the wound epithelium and normal epidermis. Staining was markedly decreased in the wound epithelium after denervation but not in the epidermis. Therefore, in both newt and axolotl the regenerating axons induce nAG protein in the nerve sheath and subsequently the protein is expressed by gland cells, under (newt) or within (axolotl) the wound epithelium, which discharge by a holocrine mechanism. These findings serve to unify the nerve dependence of limb regeneration. [source]

Cyclooxygenase-2 expression in dermatofibroma and dermatofibrosarcoma protuberans

JOURNAL OF CUTANEOUS PATHOLOGY, Issue 6 2008
Neta Adler
Background:, Dermatofibroma (DF) and dermatofibrosarcoma protuberans (DFSP) occasionally resemble each other histologically but differ in histogenesis and biological behavior. This study sought to determine if these lesions can be differentiated by the quantity or quality of expression of cyclooxygenase-2 (COX-2), an enzyme associated with both reactive and neoplastic processes. Patients and methods:, Formalin-fixed and paraffin-embedded samples from 20 DFs and 20 DFSPs were stained immunohistochemically with antibodies directed against COX-2. Staining was evaluated semiquantitatively for percentage and intensity using a three-tiered system. DFs were graded and analyzed by cellularity. Findings within the tumors were compared with fibrocyte staining in adjacent tissue. The results were analyzed. Results:, Nineteen DFs (95%) and 15 DFSPs (75%) were immunopositive for COX-2; this difference was not statistically significant. Highly cellular DFs showed more widespread (p = 0.0039; r = 0.614) and more intense (p = 0.0586; r = 0.429) staining than less cellular DFs and more prominent staining in adjacent fibroblasts (p = 0.044; r = 0.608). Conclusions:, COX-2 immunostaining does not distinguish DFs from DFSPs. However, the enzyme is expressed more widely and more intensely in more cellular, possibly younger, DFs. The prominent expression of COX-2 in DFSP may have clinical implications for treatment with COX-2 inhibitors in tumors that are not amenable to surgery. [source]

Predominant formation of heavily pigmented dermal melanocytomas resembling ,animal-type' melanomas in hepatocyte growth factor (C57BL/6 × C3H)F1 mice following neonatal UV irradiation

JOURNAL OF CUTANEOUS PATHOLOGY, Issue 9 2007
Scott R. Florell
Background:, Transgenic mice expressing hepatocyte growth factor (HGF) develop cutaneous melanocytic tumors following neonatal UV exposure. Here, we examined the histologic spectrum of UV-induced melanocytic tumors in HGF mice on a pigmented (C57BL/6 × C3H/HeN)F1 background. Methods:, Neonatally irradiated (4000 J/m2) mice were monitored for 43 weeks, and 31/34 (91%) animals developed a total of 163 melanocytic tumors. Results:, Of 54 primary tumors analyzed, most (49/54, 91%) demonstrated exclusively dermal collections of epithelioid cells with voluminous densely pigmented cytoplasm. Seven of these also demonstrated a population of spindled cells with mitoses. Several (3/54, 6%) tumors exhibited a junctional component with melanocytes present in the epidermis. Staining with PEP8 confirmed the presence of interfollicular melanocytes at the dermal-epidermal junction in neonatal skin. Conclusions:, In contrast to HGF animals on an albino (FVB) background, HGF animals on the pigmented (C57BL/6 × C3H/HeN)F1 background do not develop classic radial growth phase melanoma but rather predominantly develop dermal melanocytomas resembling the ,animal-type' melanoma occasionally seen in humans. These results demonstrate the influence of genetic background on histologic pattern of UV-induced melanomas in mice. [source]

Comparison of Seborrheic Keratoses, Inflamed Seborrheic Keratoses, and Inverted Follicular Keratoses Using P53, BCL-1, and BCL-2

JOURNAL OF CUTANEOUS PATHOLOGY, Issue 1 2005
C. Ko
While cell cycle markers have been used to differentiate benign versus malignant lesions and to classify malignant lesions, benign keratoses have not been well studied using such markers, and the relation of the cell cycle to inflammation or irritation of benign keratoses is unclear. We compared the immunohistochemical staining patterns of 10 seborrheic keratoses, 10 inflamed seborrheic keratoses, and 10 inverted follicular keratoses using antibodies to p53, bcl-1, and bcl-2. Staining with antibodies to p53 was increased in inverted follicular keratoses compared to inflamed or non-inflamed seborrheic keratoses. Bcl-1 staining was similar in all lesions. A population of bcl-2-positive dendritic cells was seen within the epidermal portion of inverted follicular keratoses. Keratinocyte bcl-2 staining was higher in seborrheic keratoses compared to the other two keratoses. Bcl-2 may be increased in seborrheic keratoses as an anti-apoptotic mechanism while increased p53 may trigger apoptosis in inverted follicular keratoses. [source]

Detection of Vacuum Infusion of Pectinmethylesterase in Strawberry by Activity Staining

JOURNAL OF FOOD SCIENCE, Issue 3 2004
P. B ANJONGSINSIRI
ABSTRACT: Strawberry halves were infused with Valencia orange pectinmethylesterase (PME) under vacuum for 15 min at room temperature. Fruits were blotted onto pectin paper and stained for activity. Activity of PME-infused fruit was about twice (0.36 unit/mL or 0.86 unit/mg protein) that of noninfused control (0.19 unit/mL) or water-infused control (0.16 unit/mL). Instron firmness values were not significantly different (P± 0.05) between noninfused and PME-infused fruit. Firmness of PME-infused fruit was about twice that of water-infused control. Water-soluble pectin, chelator soluble pectin, alkaline soluble pectin, and total pectin ranged from 8.24 to 8.70, 3.24 to 4.56, 3.77 to 5.39, and 17.86 to 26.16 mg/100 mg alcohol insoluble solid (AIS), respectively, for all treatments. [source]

Reconstructing the Sequence of Events Surrounding Body Disposition Based on Color Staining of Bone,

JOURNAL OF FORENSIC SCIENCES, Issue 5 2009
Meaghan A. Huculak H.B.Sc.
Abstract:, Literature regarding bone color is limited to determining location of primary and secondary dispositions. This research is the first to use bone color to interpret the sequence of events surrounding body disposition. Two scenarios were compared,bones buried and then exposed on the ground surface and bones exposed then buried. Forty juvenile pig humeri with minimal tissue were used in each scenario with an additional 20 controls to determine if decomposing tissue affects bone color. Munsell Color Charts were used to record bone color of surface and 2.5 cm cross-sections. Results reveal five main surface colors attributed to soil, sun, hemolysis, decomposition, and fungi. Fungi on buried bones suggests prior surface exposure. Cross-sections of strictly buried bones are identical to buried then exposed bone, stressing the importance of bone surface analysis. Cross-sectioning may help verify remains have been exposed then buried. Decomposition of excess tissue creates minimal color staining. [source]

Major muscle systems in the larval caenogastropod, Ilyanassa obsoleta, display different patterns of development

JOURNAL OF MORPHOLOGY, Issue 10 2009
Carol C.E. Evans
Abstract This study describes the anatomical and developmental aspects of muscular development from the early embryo to competent larval stage in the gastropod Ilyanassa obsoleta. Staining of F-actin revealed differential spatial and temporal patterns of several muscles. In particular, two major muscles, the larval retractor and pedal retractor muscles originate independently and display distinct developmental patterns similar to observations in other gastropod species. Additionally, together with the larval retractor muscle, the accessory larval muscle developed in the embryo at the trochophore stage. Therefore, both these muscles develop prior to ontogenetic torsion. The pedal retractor muscle marked the most abundant growth in the mid veliger stage. Also during the middle stage, the metapodial retractor muscle and opercular retractor muscle grew concurrently with development of the foot. We show evidence that juvenile muscles, such as the buccal mass muscle and siphon muscle develop initially during the late veliger stage. Collectively, these findings substantiate that larval myogenesis involves a complex sequence of events that appear evolutionary conserved within the gastropods, and set the stage for future studies using this model species to address issues concerning the evolution and eventual fates of larval musculature in molluscs. J. Morphol., 2009. © 2009 Wiley-Liss, Inc. [source]

Rho-associated kinase (ROCK) inhibitor, Y27632, promotes neurite outgrowth in PC12 cells in the absence of NGF

JOURNAL OF NEUROCHEMISTRY, Issue 2002
R. Nath
Neurite extension and retraction are very important processes in the formation of neuronal networks. A strategy for fostering axonal regrowth/regeneration of injured adult neurons is attractive therapeutically for various diseases such as traumatic brain injury, stroke and Alzheimer's disease. The Rho family of small GTPases, including Rac and Cdc42 have been shown to be involved in promoting neurite outgrowth. On the other hand, activation of RhoA induces collapse of growth cone and retraction of neurites. Rho-associated kinase (ROCK) an effector molecule of RhoA, is downstream of a number of axonal outgrowth and growth cone collapse inhibition mechanisms. In the present study, we sought to identify the role of ROCK in neurite outgrowth in PC12 cells. Y27632, a specific inhibitor of ROCK, induced a robust increase in neurite outgrowth in these cells within 24,48 h as visualized by phase contrast microscopy. Staining with FITC-tubulin or phalloidin show extended neurites in PC12 cells treated with Y27632, comparable to that with 100 ng/mL of NGF. Assessment of other biochemical markers of neurite outgrowth such as GAP43, neurofilament and tyrosine hydroxylase phosphorylation further indicates that inhibition of ROCK in PC12 cells causes differentiation of these cells to a neuronal phenotype. [source]

Antibacterial, antiviral, antiproliferative and apoptosis-inducing properties of Brackenridgea zanguebarica (Ochnaceae)

JOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 8 2006
Maren Möller
Brackenridgea zanguebarica is a small tree that is used in traditional African medicine as a type of cure-all for many diseases, including the treatment of wounds. The yellow bark of B. zanguebarica was used for the preparation of an ethanolic extract, which was tested in various concentrations against eleven bacteria, Herpes simplex virus type 1 (HSV-1) and different human tumour cell lines. The extract that contains different polyphenolic substances like calodenin B. Cell growth inhibition, assessed via MTT-assay, was found in all tested human cell lines with IC50 values (concentration of extract that reduced cell viability by 50%) between 33 ,g dry extract/mL for HL-60 human myeloid leukaemia cells and 93 ,g dry extract/mL for HaCaT human keratinocytes. Staining with Annexin-V-FLUOS and JC-1 followed by subsequent analysis via flow cytometry revealed significant apoptosis-inducing properties. Analysis of caspase activity using a fluorogenic caspase-3 substrate showed a significant caspase activity in Jurkat T-cells after incubation with the extract. The bark extract had a pronounced activity against free HSV-1 and a strong antibacterial activity against Gram-positive strains (MICs: 6,24 ,g dry extract/mL), which are often involved in skin infections. Additionally, no irritating properties of the extract could be observed in hen-egg test chorioallantoic membrane (HET-CAM) assay. These findings give a rationale for the traditional use of B. zanguebarica and are a basis for further analysis of the plant's components, their biological activity, and its use in modern phytotherapy. [source]

Riluzole prolongs survival time and alters nuclear inclusion formation in a transgenic mouse model of Huntington's disease

MOVEMENT DISORDERS, Issue 4 2002
Johannes Schiefer MD
Abstract Glutamate excitotoxicity has been suggested to contribute to the pathogenesis of Huntington's disease (HD). Riluzole is a substance with glutamate antagonistic properties that is used for neuroprotective treatment in amyotrophic lateral sclerosis and which is currently tested in clinical trials for treatment of HD. R6/2 transgenic mice, which express exon 1 of the human HD gene with an expanded CAG triplet repeat, serve as a well-characterized mouse model for HD with progressing neurological abnormalities and limited survival. We treated R6/2 HD transgenic mice with riluzole orally beginning at a presymptomatic stage until death to investigate its potential neuroprotective effects in this mouse model and found that survival time in the riluzole group was significantly increased in comparison to placebo-treated transgenic controls. Additionally, the progressive weight loss was delayed and significantly reduced by riluzole treatment; behavioral testing of motor coordination and spontaneous locomotor activity, however, showed no statistically significant differences. We also examined the formation of the HD characteristic neuronal intranuclear inclusions (NII) immunohistologically. At a late disease stage, striatal NII from riluzole-treated transgenic mice showed profound changes in ubiquitination, i.e., NII were less ubiquitinated and surrounded by ubiquitinated micro-aggregates. Staining with antibodies directed against the mutated huntingtin revealed no significant difference in this component of NII. Taken together, these data suggest that riluzole is a promising candidate for neuroprotective treatment in human HD. © 2002 Movement Disorder Society [source]

A flow cytometry-based assay for measuring invasion of red blood cells by Plasmodium falciparum,

AMERICAN JOURNAL OF HEMATOLOGY, Issue 4 2010
Amy K. Bei
Variability in the ability of the malaria parasite Plasmodium falciparum to invade human erythrocytes is postulated to be an important determinant of disease severity. Both the parasite multiplication rate and erythrocyte selectivity are important parameters that underlie such variable invasion. We have established a flow cytometry-based method for simultaneously calculating both the parasitemia and the number of multiply-infected erythrocytes. Staining with the DNA-specific dye SYBR Green I allows quantitation of parasite invasion at the ring stage of parasite development. We discuss in vitro and in vivo applications and limitations of this method in relation to the study of parasite invasion. Am. J. Hematol., 2010. © 2010 Wiley-Liss, Inc. [source]

Overexpression of p53 protein and MDM2 in papillary carcinomas of the thyroid: Correlations with clinicopathologic features

PATHOLOGY INTERNATIONAL, Issue 1 2001
Satoshi Horie
Expression of p53 protein and MDM2 was evaluated in paraffin-embedded tissue from 78 patients with papillary carcinomas of the thyroid (PCT), in order to elucidate the relationship between them and their correlations with some clinicopathologic features implicated in tumor progression. These proteins were expressed in nuclei of tumor cells, but not in non-tumor cells. Staining was defined as positive when 10% or more of tumor cells expressed these proteins. The number of cases positive for p53 protein was 21/78 (27%), and that positive for MDM2 was 26/78 (33%). Co-overexpression of p53 protein and MDM2 was observed in 12/78 cases (15%). A significant positive relationship was found between them (P < 0.01); p53-positive cases tended to be also positive for MDM2 and vice versa. Statistical analysis revealed that overexpression of p53 protein significantly correlated with large tumor size (P = 0.0271) and the presence of capsular invasion (P = 0.04). There were significant positive correlations between tumor size and intrathyroidal invasion and between tumor size and capsular invasion in PCT, suggesting that p53 protein overexpression is associated only with tumor progression (tumor size). However, we could not find any significant correlations between MDM2 expression and clinicopathologic features. Our findings suggest that overexpression of p53 protein and MDM2 in papillary carcinoma of the thyroid is associated with the progression of the tumors, and that p53 may be a marker of the progression of PCT. [source]

Effects of temperature of adults and eggs on the induction of embryonic diapause in the band-legged ground cricket, Dianemobius nigrofasciatus

PHYSIOLOGICAL ENTOMOLOGY, Issue 3 2006
EIJI FUKUMOTO
Abstract Eggs laid by adult female Dianemobius nigrofasciatus, reared under long-day (LD 16 : 8 h, 25 °C) or short-day (LD 12 : 12 h, 25 °C) conditions from the nymphal stage, are kept at several constant temperatures. At 22.5,30.0 °C, eggs laid by long-day adults show lower incidences of diapause than those laid by short-day adults. In both eggs laid by adults under long-day conditions and those under short-day conditions, the higher the temperature at which the eggs are kept, the lower the incidence of diapause. When eggs of long-day adults are exposed to a low-temperature pulse (10 °C, 24 h) on the day of deposition (day 0), the incidence of diapause increases. The low-temperature pulse on day 1 does not increase the incidence of diapause. By contrast, when the eggs of short-day adults are exposed to a high-temperature pulse (35 °C, 24 h) on day 0 or day 1, the incidence of diapause decreases. The temperature pulses on day 0 are more effective at diapause prevention. Staining of diapause eggs by the Feulgen,Rossenbeck method shows that the eggs enter diapause at the blastoderm stage, which is on day 1 or day 2 at 25 °C. The exposure of adults to long days and higher temperatures prevents the eggs from entering diapause. In D. nigrofasciatus, embryonic diapause is controlled by maternal effects, adult photoperiod and temperature, and egg temperature before or at diapause. [source]

Inhibitory effects of furanone metabolites of a rhizobacterium, Pseudomonas jessenii, on phytopathogenic Aphanomyces cochlioides and Pythium aphanidermatum

PLANT PATHOLOGY, Issue 1 2010
A. 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]

Inhibition of the development of leaf rust (Puccinia recondita) by treatment of wheat with allopurinol and production of a hypersensitive-like reaction in a compatible host

PLANT PATHOLOGY, Issue 3 2000
A. L. Ádám
The effect of allopurinol [4-hydroxypyrazolo (3,4- d) pyrimidine], a purine analogue inhibitor of xanthine oxidase (XO) enzyme, was studied in the host,pathogen combination of Triticum aestivum,Puccinia recondita f.sp. tritici. Analysis of purines and pyrimidines in the allopurinol-treated wheat seedlings showed marked accumulation of xanthine, suggesting the inplanta inhibition of XO activity. In the incompatible wheat,rust interaction application of allopurinol as a drench, even at the highest concentration (50 ,m), did not change the hypersensitive reaction phenotype; only the number of lesions was slightly reduced. Allopurinol treatment decreased the augmented rate of electrolyte leakage and lipid peroxidation associated with the hypersensitive response (HR), an effect probably related to the inhibition of rust development by allopurinol. By contrast, in the case of the compatible wheat,leaf-rust combination the reaction type was strongly affected. The formation of uredia and production of uredospores were diminished or completely inhibited depending on the concentration of allopurinol, which was applied either as a drench (3.125,50 ,m) or as a foliar spray (100,400 ,m) to plants grown in perlite. At the highest allopurinol concentration in the drench, the compatible reaction type changed to a hypersensitive-like necrotic reaction. Significant increases in electrolyte leakage and lipid peroxidation (characteristic of the HR) were found 4,6 days after infection in susceptible plants treated with allopurinol. Staining of leaf slices from allopurinol-treated and compatible rust-infected plants with Evans blue indicated cell death surrounding the pustules, while at this stage no cell death was detected in infected leaves without allopurinol treatment. The above results suggest that XO is not the main source of the generation of active oxygen species in wheat during the HR to leaf rust. [source]

1141478081 Cleavage of integrin by calpain in patients with recurrent miscarriage

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 6 2006
K Nozawa
Problem:, In our previous study, we have been reported that calpain, a calcium-dependent cysteine protease, activated by increasing intracellular calcium-ion concentration in endometrial cell, cause endometrial dysfunction for implantation. In this study, we investigated the existence and contribution of calpains, their endogenous inhibitor calpastatin, and their substrate such as integrin ,3 and ,-fodrin in deciduas of patients with recurrent miscarriage. Method of Study:, Deciduas were surgically collected from 31 patients with recurrent miscarriage and 23 healthy women with informed consent. Immunohistochemistry, SDS-PAGE and western blot analysis were performed using antibodies against calpain, calpastatin, integrin ,3 and ,-fodrin. Results:, Staining of ,-calpain, m-calpain, capastatin, integrin ,3 and ,-fodrin were observed in the cytoplasm of stromal and epithelial cells in deciduas using immunohistochemistry. No significant differences were observed in staining patterns. Western blot analysis shows no significant differences in m-calpain, calpastatin and ,-fodrin, while ,-calpain was significantly higher and integrin ,3 was lower in subject. Conclusions:, The result provided that cleavage of integrin ,3 by ,-calpain may cause adverse effect in the mechanism of early pregnancy. [source]

Pancreas Allograft Biopsies with Positive C4d Staining and Anti-Donor Antibodies Related to Worse Outcome for Patients

AMERICAN JOURNAL OF TRANSPLANTATION, Issue 7 2010
H. De Kort
C4d+ antibody-mediated rejection following pancreas transplantation has not been well characterized. Therefore, we assessed the outcomes of 27 pancreas transplantation patients (28 biopsies), with both C4d staining and donor-specific antibodies (DSA) determined, from a cohort of 257 patients. The median follow-up was 50 (interquartile range [IQR] 8,118) months. Patients were categorized into 3 groups: group 1, patients with minimal or no C4d staining and no DSA (n = 13); group 2, patients with either DSA present but no C4d, diffuse C4d+ and no DSA or focal C4d+ and DSA (n = 6); group 3, patients with diffuse C4d+ staining and DSA (n = 9). Active septal inflammation, acinar inflammation and acinar cell injury/necrosis were significantly more abundant in group 3 than in group 2 (respective p-values: 0.009; 0.033; 0.025) and in group 1 (respective p-values: 0.034; 0.009; 0.002). The overall uncensored pancreas graft survival rate for groups 1, 2 and 3 were 53.3%, 66.7% and 34.6%, respectively (p = 0.044). In conclusion, recipients of pancreas transplants with no C4d or DSA had excellent long-term graft survival in comparison with patients with both C4d+ and DSA present. Hence, C4d should be used as an additional marker in combination with DSA in the evaluation of pancreas transplant biopsies. [source]

Endothelial Gene Expression in Kidney Transplants with Alloantibody Indicates Antibody-Mediated Damage Despite Lack of C4d Staining

AMERICAN JOURNAL OF TRANSPLANTATION, Issue 10 2009
Banu Sis
Anti-HLA alloantibody is a risk factor for graft loss, but does not indicate which kidneys are experiencing antibody-mediated rejection (ABMR). C4d staining in biopsies is specific for ABMR but insensitive. We hypothesized that altered expression of endothelial genes due to alloantibody acting on the microcirculation would be sensitive indicator of ABMR. We identified 119 endothelial-associated transcripts (ENDATs) from literature, and studied their expression by microarrays in 173 renal allograft biopsies for cause. Mean ENDAT expression was increased in all rejection but was higher in ABMR than in T-cell-mediated rejection and correlated with histopathologic lesions of ABMR, and alloantibody. Many individual ENDATs were increased in ABMR and predicted graft loss. Kidneys with high ENDATs and antibody showed increased lesions of ABMR and worse prognosis in comparison to controls. Only 40% of kidneys with high ENDAT expression and chronic ABMR or graft loss were diagnosed by C4d positivity. High ENDAT expression with antibody predicts graft loss with higher sensitivity (77% vs. 31%) and slightly lower specificity (71% vs. 94%) than C4d. The results were validated in independent set of 82 kidneys. High renal endothelial transcript expression in patients with alloantibody is indicator of active antibody-mediated allograft damage and poor graft outcome. [source]

C4d and C3d Staining in Biopsies of ABO- and HLA-Incompatible Renal Allografts: Correlation with Histologic Findings

AMERICAN JOURNAL OF TRANSPLANTATION, Issue 8 2006
M. Haas
Biopsies of ABO-incompatible and positive crossmatch (HLA-incompatible) renal allografts were retrospectively examined to compare results of C4d and C3d staining, and the correlation between such staining and histologic findings suggestive of antibody-mediated rejection (AMR). A total of 75 biopsies (55 protocol, 17 for graft dysfunction, 3 for other indications) of 24 ABO-incompatible grafts and 244 biopsies (103 protocol, 129 for graft dysfunction, 12 for other indications) of 66 HLA-incompatible grafts were examined; all were stained for C4d and ,40% for C3d. In ABO-incompatible grafts, 80% of protocol biopsies and 59% performed for graft dysfunction showed C4d staining in peritubular capillaries (PTC); this staining was not correlated with neutrophil margination in PTC. In HLA-incompatible grafts, PTC C4d was present in 26% of protocol biopsies and 60% of biopsies for graft dysfunction; 92% of biopsies with >1+ (0,4+ scale), diffuse PTC C4d had ,1+ margination and/or thrombotic microangiopathy (TMA), compared with 12% of C4d-negative biopsies. C3d was somewhat more predictive of margination than C4d in ABO-incompatible, but not HLA-incompatible, grafts. In summary, while PTC C4d deposition indicates probable AMR in biopsies of HLA-incompatible grafts, including protocol biopsies, there is no histologic evidence that C4d deposition is correlated with injury in most ABO-incompatible grafts. [source]

Direct Polymerase Synthesis of Reactive Aldehyde-Functionalized DNA and Its Conjugation and Staining with Hydrazines,

ANGEWANDTE CHEMIE, Issue 6 2010
Veronika Raindlová
In zwei Stufen zu reaktiver Aldehyd-modifizierter DNA: durch Suzuki-Kreuzkupplung halogenierter Nucleosidtriphosphate (dNTPs) mit 4-Formylthiophen-2-boronsäure und Polymerase- vermittelten Einbau der modifizierten Nucleotide in DNA (siehe Schema; PEX=Primerverlängerung, PCR=Polymerasekettenreaktion). Die Bildung von Hydrazonen mit Arylhydrazinen unter wässrigen Bedingungen wurde zum Anfärben der DNA verwendet. [source]

Anti-angiogenic factor endostatin in osteosarcoma

APMIS, Issue 10 2009
HYUN-SOO KIM
Neoplastic neovascularization is regulated not only by stimulators, but also by inhibitors of angiogenesis and might be the result of a net balance between the positive and negative regulators. Endostatin (ES) is a potent inhibitor of angiogenesis. The expression of ES has not been investigated in patients with osteosarcomas (OSAs). The aim of this study was to determine whether there is a correlation between the expression of ES and clinicopathologic parameters and/or outcomes in patients with OSAs. We made tissue microarrays from 46 cases of OSA and analyzed the expression of ES using immunohistochemistry. Staining was assessed in a semi-quantitative manner by scoring the proportion of positive tumor cells over the total number of tumor cells. A sample was defined as ES-positive when 10% or more of the tumor cells were stained positively throughout the tumor core. ES was localized to the cytoplasm of the tumor cells. 32.6% (15/46) of the patients were ES-positive. The expression of ES was positively correlated with tumor size (p = 0.011), histologic grade (p = 0.034), stage (p = 0.025), and distant metastasis (p = 0.036). Our results suggest that the expression of ES is increased in OSA, and ES may be used as a prognostic marker in patients with OSAs. [source]

A novel model of wound healing in the SCID mouse using a cultured human skin substitute

AUSTRALASIAN JOURNAL OF DERMATOLOGY, Issue 1 2009
Martin L Windsor
SUMMARY Studies of skin graft behaviour in rodent excisional wound models are limited by the dominance of wound contracture and graft sloughing as primary healing responses. To slow skin contraction, polytetrafluoroethylene (Teflon) rings were inserted into dorso-lateral full-thickness wounds in SCID mice. Cultured skin substitutes (OrCel), composed of cultured human keratinocytes and fibroblasts in a bovine collagen sponge, were implanted within the rings. Examination and histology of grafts 14 days later showed graft take in four of six recipients, with 90% epithelialization and wound contraction of 31,47%. Immunohistochemical studies, using human-specific antisera to distinguish graft from host tissues, showed that regenerated tissue was predominantly human. Staining with anticytokeratin, revealed a multilayered, stratified neoepidermis. HBG were identified in keratinocytes in all epidermal layers. Langerhans cells were absent. Antihuman vimentin, used as a fibroblast marker, confirmed that cells of the neodermis were primarily of human origin. Neoepidermal keratinocytes, primarily in the basal and suprabasal layers, were also stained. Results suggest that the poly(tetrafluoroethylene) ring inhibited graft sloughing and provided a more favourable environment for the skin substitute to regenerate a substantially normal human skin. [source]

Morphological and immunohistochemical studies on cleft palates induced by 2,3,7,8-tetrachlorodibenzo- p -dioxin in mice

CONGENITAL ANOMALIES, Issue 2 2008
Kumiko Fujiwara
ABSTRACT Morphological and immunohistological examinations were performed to reveal the mechanisms of cleft palate induction by 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD). ICR strain mice 8,10 weeks of age were used in the study. TCDD was administered in olive oil on gestation day (GD) 12.5 with gastric tubes at 40 ,g/kg. From GD 13.5 to 16.5, palates were examined by scanning electron microscopy (SEM), hematoxyline,eosin (HE) staining, and immunohistochemical staining of FGFR1/2, TGF-,3, MSX1 and LHX8. In the control group, both of the palatal shelves began elevating on GD 14.0 and finished within 6 h. After the elevation, all of the shelves had completely fused with each other on GD 14.5. In the TCDD-treated group, palatal shelves elevated 1 day later than in the control group. However, all palates had elevated by GD 15.0. After the elevation, the shelves contacted each other and fused; however, they were separated on GD16.0. HE staining showed that medial edge epithelium (MEE) was thinner in the TCDD group than in the control group. MEE observed under a high magnification (×2500) exhibited filopodia-like filaments and the cells were bulged in the control group. In contrast, in the TCDD group, no filaments were observed and the cells were flat with unclear boundaries. Immunohistologically, there were no characteristic findings except for FGFR1. FGFR1 was not expressed in the TCDD group after the fusion phase (GD 14.5). TCDD induces many morphological and molecular changes to MEE cells and causes cleft palates. [source]