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Golshani, Maryam, Waheed Ur Rahman, Adriana Osickova, Jana Holubova, Jinery Lora, Nataliya Balashova, Peter Sebo, and Radim Osicka. "Filamentous Hemagglutinin of Bordetella pertussis Does Not Interact with the β2 Integrin CD11b/CD18." International Journal of Molecular Sciences 23, no. 20 (October 20, 2022): 12598. http://dx.doi.org/10.3390/ijms232012598.
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The pertussis agent Bordetella pertussis produces a number of virulence factors, of which the filamentous hemagglutinin (FhaB) plays a role in B. pertussis adhesion to epithelial and phagocytic cells. Moreover, FhaB was recently found to play a crucial role in nasal cavity infection and B. pertussis transmission to new hosts. The 367 kDa FhaB protein translocates through an FhaC pore to the outer bacterial surface and is eventually processed to a ~220 kDa N-terminal FHA fragment by the SphB1 protease. A fraction of the mature FHA then remains associated with bacterial cell surface, while most of FHA is shed into the bacterial environment. Previously reported indirect evidence suggested that FHA, or its precursor FhaB, may bind the β2 integrin CD11b/CD18 of human macrophages. Therefore, we assessed FHA binding to various cells producing or lacking the integrin and show that purified mature FHA does not bind CD11b/CD18. Further results then revealed that the adhesion of B. pertussis to cells does not involve an interaction between the bacterial surface-associated FhaB and/or mature FHA and the β2 integrin CD11b/CD18. In contrast, FHA binding was strongly inhibited at micromolar concentrations of heparin, corroborating that the cell binding of FHA is ruled by the interaction of its heparin-binding domain with sulfated glycosaminoglycans on the cell surface.
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Julio, Steven M., and Peggy A. Cotter. "Characterization of the Filamentous Hemagglutinin-Like Protein FhaS in Bordetella bronchiseptica." Infection and Immunity 73, no. 8 (August 2005): 4960–71. http://dx.doi.org/10.1128/iai.73.8.4960-4971.2005.
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ABSTRACT Filamentous hemagglutinin (FHA) is a large (>200 kDa), rod-shaped protein expressed by bordetellae that is both surface-associated and secreted. FHA mediates bacterial adherence to epithelial cells and macrophages in vitro and is absolutely required for tracheal colonization in vivo. The recently sequenced Bordetella bronchiseptica genome revealed the presence of a gene, fhaS, that is nearly identical to fhaB, the FHA structural gene. We show that although fhaS expression requires the BvgAS virulence control system, it is maximal only under a subset of conditions in which BvgAS is active, suggesting an additional level of regulation. We also show that, like FHA, FhaS undergoes a C-terminal proteolytic processing event and is both surface-associated and secreted and that export across the outer membrane requires the channel-forming protein FhaC. Unlike FHA, however, FhaS was unable to mediate adherence of B. bronchiseptica to epithelial cell lines in vitro and was not required for respiratory tract colonization in vivo. In a coinfection experiment, a ΔfhaS strain was out-competed by wild-type B. bronchiseptica, indicating that fhaS is expressed in vivo and that FhaS contributes to bacterial fitness in a manner revealed when the mutant must compete with wild-type bacteria. These data suggest that FHA and FhaS perform distinct functions during the Bordetella infectious cycle. A survey of various Bordetella strains revealed two distinct fhaS alleles that segregate according to pathogen host range and that B. parapertussis hu most likely acquired its fhaS allele from B. pertussis horizontally, suggesting fhaS may contribute to host-species specificity.
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Tsai, Ming-Daw. "FHA." Structure 10, no. 7 (July 2002): 887–88. http://dx.doi.org/10.1016/s0969-2126(02)00795-5.
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Wilson, Dan R., Annette Siebers, and B. Brett Finlay. "Antigenic Analysis of Bordetella pertussis Filamentous Hemagglutinin with Phage Display Libraries and Rabbit Anti-Filamentous Hemagglutinin Polyclonal Antibodies." Infection and Immunity 66, no. 10 (October 1, 1998): 4884–94. http://dx.doi.org/10.1128/iai.66.10.4884-4894.1998.
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ABSTRACT Although substantial advancements have been made in the development of efficacious acellular vaccines against Bordetella pertussis, continued progress requires better understanding of the antigenic makeup of B. pertussis virulence factors, including filamentous hemagglutinin (FHA). To identify antigenic regions of FHA, phage display libraries constructed by using random fragments of the 10-kbp EcoRI fragment ofB. pertussis fhaB were affinity selected with rabbit anti-FHA polyclonal antibodies. Characterization of antibody-reactive clones displaying FHA-derived peptides identified 14 antigenic regions, each containing one or more epitopes. A number of clones mapped within regions containing known or putative FHA adhesin domains and may be relevant for the generation of protective antibodies. The immunogenic potential of the phage-displayed peptides was assessed indirectly by comparing their recognition by antibodies elicited by sodium dodecyl sulfate (SDS)-denatured and native FHA and by measuring the inhibition of this recognition by purified FHA. FHA residues 1929 to 2019 may contain the most dominant linear epitope of FHA. Clones mapping to this region accounted for ca. 20% of clones recovered from the initial library selection and screening procedures. They are strongly recognized by sera against both SDS-denatured and native FHA, and this recognition is readily inhibited by purified FHA. Given also that this region includes a factor X homolog (J. Sandros and E. Tuomanen, Trends Microbiol. 1:192–196, 1993) and that the single FHA epitope (residues 2001 to 2015) was unequivocally defined in a comparable study by E. Leininger et al. (J. Infect. Dis. 175:1423–1431, 1997), peptides derived from residues of 1929 to 2019 of FHA are strong candidates for future protection studies.
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Geisthövel. "Functional hyperandrogenism – classification, etiology, diagnostic and therapy." Therapeutische Umschau 59, no. 4 (April 1, 2002): 163–73. http://dx.doi.org/10.1024/0040-5930.59.4.163.
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Die dargestellte Klassifizierung des Funktionellen Hyperandrogenismus (FHA) basiert auf lange und seit jüngerer Zeit bekannten klinischen Erkenntnissen, die durch neuere molekularbiologische Vorstellungen untermauert werden. Man kann annehmen, dass sich der FHA aus verschiedenen, organ- oder systemspezifischen Entitäten zusammensetzt, die letztlich unterschiedliche Diagnostik- und Therapiestrategien zur Folge haben. Die Bezeichnung PCOS», welche die verschiedenen Entitäten ungenau und schieflastig umfasst, sollte durch die entsprechenden, adäquaten Begriffsbestimmungen ersetzt werden. Obwohl schon umfangreiche Denkansätze und Kenntnisse zu molekularbiologischen Abläufen beim FHA vorliegen, sind mit Ausnahme der hier nicht besprochenen FHA-Gruppe III (FAHA) die ätiologischen Folgerungen noch nicht gefestigt und die diagnostischen wie therapeutischen Konsequenzen weiterhin limitiert. Allerdings sind in der nahen Zukunft auf der Basis der jüngsten Erkenntnisse des menschlichen Genoms und den neuen genetischen screenings-Möglichkeiten (Microarrays) Weiterentwicklungen zu erwarten.
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Durocher, Daniel, and Stephen P. Jackson. "The FHA domain." FEBS Letters 513, no. 1 (December 20, 2001): 58–66. http://dx.doi.org/10.1016/s0014-5793(01)03294-x.
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Alonso, Sylvie, Kévin Pethe, Nathalie Mielcarek, Dominique Raze, and Camille Locht. "Role of ADP-Ribosyltransferase Activity of Pertussis Toxin in Toxin-Adhesin Redundancy with Filamentous Hemagglutinin duringBordetella pertussis Infection." Infection and Immunity 69, no. 10 (October 1, 2001): 6038–43. http://dx.doi.org/10.1128/iai.69.10.6038-6043.2001.
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ABSTRACT Pertussis toxin (PT) and filamentous hemagglutinin (FHA) are two major virulence factors of Bordetella pertussis. FHA is the main adhesin, whereas PT is a toxin with an A-B structure, in which the A protomer expresses ADP-ribosyltransferase activity and the B moiety is responsible for binding to the target cells. Here, we show redundancy of FHA and PT during infection. Whereas PT-deficient and FHA-deficient mutants colonized the mouse respiratory tract nearly as efficiently as did the isogenic parent strain, a mutant deficient for both factors colonized substantially less well. This was not due to redundant functions of PT and FHA as adhesins, since in vitro studies of epithelial cells and macrophages indicated that FHA, but not PT, acts as an adhesin. An FHA-deficient B. pertussis strain producing enzymatically inactive PT colonized as poorly as did the FHA-deficient, PT-deficient strain, indicating that the ADP-ribosyltransferase activity of PT is required for redundancy with FHA. Only strains producing active PT induced a local transient release of tumor necrosis factor alpha (TNF-α), suggesting that the pharmacological effects of PT are the basis of the redundancy with FHA, through the release of TNF-α. This may lead to damage of the pulmonary epithelium, allowing the bacteria to colonize even in the absence of FHA.
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Poulain-Godefroy, Odile, Nathalie Mielcarek, Nathalie Ivanoff, Franck Remoué, Anne-Marie Schacht, Nigel Phillips, Camille Locht, André Capron, and Gilles Riveau. "Bordetella pertussis Filamentous Hemagglutinin Enhances the Immunogenicity of Liposome-Delivered Antigen Administered Intranasally." Infection and Immunity 66, no. 4 (April 1, 1998): 1764–67. http://dx.doi.org/10.1128/iai.66.4.1764-1767.1998.
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ABSTRACT In an attempt to increase the immunogenicity of mucosally delivered antigens, we incorporated the Bordetella pertussisfilamentous hemagglutinin (FHA) adhesin into liposomes containing the glutathione S-transferase of Schistosoma mansoni (Sm28GST) as a model antigen. Outbred mice immunized twice intranasally with liposomes containing a constant suboptimal dose of Sm28GST and increasing doses of FHA produced anti-Sm28GST antibodies in a FHA dose-dependent manner. The addition of 3 μg of FHA to the liposomes induced more than 10-fold-higher anti-Sm28GST antibody titers, compared to those induced by liposomes without FHA. The presence of FHA did not alter the nature of the humoral immune response, and the sera contained anti-Sm28GST immunoglobulin G1 (IgG1), IgG2a, and IgG2b. However, anti-Sm28GST IgA was only detected when at least 3 μg of FHA was added to the preparation. These results show a promising potential for FHA to enhance the immunogenicity of mucosally administered antigens incorporated into liposomes.
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Luo, Shukun, Xiaoran Xin, Li-Lin Du, Keqiong Ye, and Yi Wei. "Dimerization Mediated by a Divergent Forkhead-associated Domain Is Essential for the DNA Damage and Spindle Functions of Fission Yeast Mdb1." Journal of Biological Chemistry 290, no. 34 (July 9, 2015): 21054–66. http://dx.doi.org/10.1074/jbc.m115.642538.
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MDC1 is a key factor of DNA damage response in mammalian cells. It possesses two phospho-binding domains. In its C terminus, a tandem BRCA1 C-terminal domain binds phosphorylated histone H2AX, and in its N terminus, a forkhead-associated (FHA) domain mediates a phosphorylation-enhanced homodimerization. The FHA domain of the Drosophila homolog of MDC1, MU2, also forms a homodimer but utilizes a different dimer interface. The functional importance of the dimerization of MDC1 family proteins is uncertain. In the fission yeast Schizosaccharomyces pombe, a protein sharing homology with MDC1 in the tandem BRCA1 C-terminal domain, Mdb1, regulates DNA damage response and mitotic spindle functions. Here, we report the crystal structure of the N-terminal 91 amino acids of Mdb1. Despite a lack of obvious sequence conservation to the FHA domain of MDC1, this region of Mdb1 adopts an FHA-like fold and is therefore termed Mdb1-FHA. Unlike canonical FHA domains, Mdb1-FHA lacks all the conserved phospho-binding residues. It forms a stable homodimer through an interface distinct from those of MDC1 and MU2. Mdb1-FHA is important for the localization of Mdb1 to DNA damage sites and the spindle midzone, contributes to the roles of Mdb1 in cellular responses to genotoxins and an antimicrotubule drug, and promotes in vitro binding of Mdb1 to a phospho-H2A peptide. The defects caused by the loss of Mdb1-FHA can be rescued by fusion with either of two heterologous dimerization domains, suggesting that the main function of Mdb1-FHA is mediating dimerization. Our data support that FHA-mediated dimerization is conserved for MDC1 family proteins.
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Li, J., G. I. Lee, S. R. Van Doren, and J. C. Walker. "The FHA domain mediates phosphoprotein interactions." Journal of Cell Science 113, no. 23 (December 1, 2000): 4143–49. http://dx.doi.org/10.1242/jcs.113.23.4143.
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The forkhead-associated (FHA) domain is a phosphopeptide-binding domain first identified in a group of forkhead transcription factors but is present in a wide variety of proteins from both prokaryotes and eukaryotes. In yeast and human, many proteins containing an FHA domain are found in the nucleus and involved in DNA repair, cell cycle arrest, or pre-mRNA processing. In plants, the FHA domain is part of a protein that is localized to the plasma membrane and participates in the regulation of receptor-like protein kinase signaling pathways. Recent studies show that a functional FHA domain consists of 120–140 amino acid residues, which is significantly larger than the sequence motif first described. Although FHA domains do not exhibit extensive sequence similarity, they share similar secondary and tertiary structures, featuring a sandwich of two anti-parallel (beta)-sheets. One intriguing finding is that FHA domains may bind phosphothreonine, phosphoserine and sometimes phosphotyrosine, distinguishing them from other well-studied phosphoprotein-binding domains. The diversity of proteins containing FHA domains and potential differences in binding specificities suggest the FHA domain is involved in coordinating diverse cellular processes.
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Wang, Si-Han, Shiao-Pieng Lee, Chung-Wei Yang, and Chun-Min Lo. "Surface Modification of Biodegradable Mg-Based Scaffolds for Human Mesenchymal Stem Cell Proliferation and Osteogenic Differentiation." Materials 14, no. 2 (January 18, 2021): 441. http://dx.doi.org/10.3390/ma14020441.
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Magnesium alloys with coatings have the potential to be used for bone substitute alternatives since their mechanical properties are close to those of human bone. However, the surface modification of magnesium alloys to increase the surface biocompatibility and reduce the degradation rate remains a challenge. Here, FHA-Mg scaffolds were made of magnesium alloys and coated with fluorohydroxyapatite (FHA). Human mesenchymal stem cells (hMSCs) were cultured on FHA-Mg scaffolds and cell viability, proliferation, and osteogenic differentiation were investigated. The results showed that FHA-Mg scaffolds display a nano-scaled needle-like structure of aggregated crystallites on their surface. The average Mg2+ concentration in the conditioned media collected from FHA-Mg scaffolds (5.8–7.6 mM) is much lower than those collected from uncoated, Mg(OH)2-coated, and hydroxyapatite (HA)-coated samples (32.1, 17.7, and 21.1 mM, respectively). In addition, compared with hMSCs cultured on a culture dish, cells cultured on FHA-Mg scaffolds demonstrated better proliferation and comparable osteogenic differentiation. To eliminate the effect of osteogenic induction medium, hMSCs were cultured on FHA-Mg scaffolds in culture medium and an approximate 66% increase in osteogenic differentiation was observed three weeks later, indicating a significant effect of the nanostructured surface of FHA-Mg scaffolds on hMSC behaviors. With controllable Mg2+ release and favorable mechanical properties, porous FHA-Mg scaffolds have a great potential in cell-based bone regeneration.
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Abramson, Tzvia, Hassya Kedem, and David A. Relman. "Proinflammatory and Proapoptotic Activities Associated with Bordetella pertussis Filamentous Hemagglutinin." Infection and Immunity 69, no. 4 (April 1, 2001): 2650–58. http://dx.doi.org/10.1128/iai.69.4.2650-2658.2001.
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ABSTRACT Filamentous hemagglutinin (FHA) is a dominant cell surface-associated Bordetella pertussis adhesin. Recognition that this protein is secreted in significant amounts and that bacterial adhesins may have other actvities, prompted an assessment of FHA effects on human macrophages. Incubation of human macrophage-like U937 cells with preparations of FHA resulted in dose-dependent cytotoxicity, with death of 95% of treated cells after 24 h. Based on the use of four independent methods, death of these cells could be largely attributed to apoptosis. FHA-associated apoptosis was also observed in THP-1 macrophage-like cells, fresh human peripheral blood monocyte-derived macrophages (MDM), and BEAS-2B human bronchial epithelial cells. Infection of MDM with wild-type B. pertussis resulted in apoptosis within 6 h, while infection with an FHA-deficient derivative strain was only 50% as effective. FHA-associated cytotoxicity was preceded by host cell secretion of tumor necrosis factor alpha (TNF-α), a potential proapoptotic factor. However, pretreatment of cells with a neutralizing anti-TNF-α monoclonal antibody inhibited only 16% of the FHA-associated apoptosis. On the other hand, a blocking monoclonal antibody directed against TNF-α receptor 1 inhibited FHA-associated apoptosis by 47.7% (P = 0.0001), suggesting that this receptor may play a role in the death pathway activated by FHA. Our in vitro data indicate that secreted and cell-associated FHA elicits proinflammatory and proapoptotic responses in human monocyte-like cells, MDM, and bronchial epithelial cells and suggest a previously unrecognized role for this prominent virulence factor in the B. pertussis-host interaction.
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Wang, Liping, Ming Wang, Yunli Wang, Yiran Shao, and Yingchun Zhu. "Facile synthesis and the phase transition mechanism of fluoridated hydroxyapatite with a hierarchical architecture." CrystEngComm 19, no. 48 (2017): 7298–306. http://dx.doi.org/10.1039/c7ce01871a.
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Mobberley-Schuman, Paula S., and Alison A. Weiss. "Influence of CR3 (CD11b/CD18) Expression on Phagocytosis of Bordetella pertussis by Human Neutrophils." Infection and Immunity 73, no. 11 (November 2005): 7317–23. http://dx.doi.org/10.1128/iai.73.11.7317-7323.2005.
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ABSTRACT CR3 (CD11b/CD18) is expressed on neutrophils, and the engagement of CR3 can promote phagocytosis. CR3 serves as the receptor for the Bordetella pertussis adhesin filamentous hemagglutinin (FHA) and for the adenylate cyclase toxin (ACT), which blocks neutrophil function. The influence of CR3, FHA, and ACT on the phagocytosis of B. pertussis by human neutrophils was examined. The surface expression and function of CR3 are regulated. Tumor necrosis factor alpha (TNF-α) and gamma interferon (IFN-γ) increased CR3 surface expression, but only TNF-α increased the ability of neutrophils to phagocytose B. pertussis, suggesting that elevated CR3 expression alone is not sufficient to promote phagocytosis. Purified FHA and pertussis toxin also increased the surface expression of CR3 on neutrophils, while ACT and the B subunit of pertussis toxin did not affect CR3 expression. FHA-mediated attachment to CR3 can lead to phagocytosis, especially in the absence of ACT. FHA mutants failed to attach and were not phagocytosed by neutrophils. Similarly, an antibody to CR3 blocked both attachment and phagocytosis. The addition of exogenous FHA enhanced the attachment and phagocytosis of wild-type B. pertussis and FHA mutants. Mutants lacking the SphB1 protease, which cleaves FHA and allows the release of FHA from the bacterial surface, were phagocytosed more efficiently than wild-type bacteria. ACT mutants were efficiently phagocytosed, but wild-type B. pertussis or ACT mutants plus exogenous ACT resisted phagocytosis. These studies suggest that the activation and surface expression of CR3, FHA expression, and the efficiency of ACT internalization all influence whether B. pertussis will be phagocytosed and ultimately killed by neutrophils.
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Amruthaluru, Saikiran, Siva Kumar Mamidi, Manu Harilal, Hariprasad Sampatirao, and Rameshbabu Nagumothu. "Synthesis of Carbonate and Fluorine Substituted Nanocrystalline Hydroxyapatite by Mechanochemical Method." Key Engineering Materials 833 (March 2020): 204–8. http://dx.doi.org/10.4028/www.scientific.net/kem.833.204.
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The present work is aimed at the synthesis of fluorine substituted and carbonate substituted hydroxyapatites (FHA, CHA) by the mechanochemical method. The shortest milling time required for the synthesis of FHA and CHA using calcium hydroxide and diammonium hydrogen phosphate as precursors was estimated. In addition to the Ca and P precursors, ammonium carbonate and ammonium fluoride were used for carbonate and fluorine substitutions, respectively. Thermal stability of the synthesized FHA and CHA was evaluated. The phase composition and crystallite size were evaluated by the X-Ray Diffraction (XRD). Fourier Transform Infrared Spectroscopy (FTIR) technique was employed to confirm the functional groups corresponding to the FHA and CHA. Thermal stability of the FHA and CHA was determined by the XRD and FTIR studies on the FHA and CHA powders annealed at 900 °C. From the XRD and FTIR results, it is observed that the 30 min milling time is the shortest time for the complete formation of FHA and CHA. The powders synthesized with a minimum milling time of 30 min exhibited better thermal stability.
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Cheng, Kui. "Effect of Mn on Protein Adsorption on Mn-TCP/FHA Coatings." Key Engineering Materials 330-332 (February 2007): 881–84. http://dx.doi.org/10.4028/www.scientific.net/kem.330-332.881.
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The effect of Mn on the protein adsorption of calcium phosphate coatings is investigated in this work. Mn containing β-tricalcium phosphate (Mn-TCP) is first prepared through a coprecipitation based methods. Then Mn-TCP is dispersed into the fluoridated hydroxyapatite (FHA) precursor sol. Mn-TCP/FHA biphasic coatings are prepared with these sols. After cell culture, it is found the amount of protein adsorbed on the coatings following this rule: Mn-TCP/FHA > β-TCP/FHA > FHA. That is ascribed to two reasons: a) Ca and PO4 releasing can promote the activity of cells; b) Mn releasing promotes protein activation even at quite low concentration.
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Bairey Merz, C. Noel, Sarah Berga, Galen Cook-Weins, Margareta Pisarska, Prediman Krishan Shah, and Chrisandra Shufelt. "OR19-6 Functional Hypothalamic Amenorrhea and Preclinical Cardiovascular Disease." Journal of the Endocrine Society 6, Supplement_1 (November 1, 2022): A249. http://dx.doi.org/10.1210/jendso/bvac150.512.
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Abstract Background Functional Hypothalamic Amenorrhea (FHA) is characterized by suppression of central GnRH drive with secondary anovulation and hypoestrogenemia, hypercortisolism, and low leptin that results from a combination of psychosocial stress and metabolic imbalance due to excess physical activity and/or weight loss. Cardiovascular disease (CVD) is the leading killer of women at all ages and CVD mortality rates are unchanged or increasing in young women aged 35-44 years. We investigated relations between FHA and preclinical CVD measured by endothelial dysfunction in premenopausal women with and without FHA, and recently menopausal women not on hormone therapy. Methods We studied 31 women with FHA, 29 eumenorrheic controls, and 30 recently menopausal women not on hormone therapy. FHA was defined as amenorrhea ≥3 consecutive months, estradiol <50 pg/mL, FSH <10 mIU/L, and LH <10 mIU/L, and exclusion of other etiologies including polycystic ovary syndrome, prolactinoma, thyroid dysfunction, and pregnancy. Eumenorrheic controls had self-reported monthly menstrual cycles, were not on hormones, with a day 22-24 progesterone level >3 ng/ml to confirm ovulation. Preclinical CVD testing for eumenorrheic controls took place days 3-5 of menses. Recently menopausal women had natural menopause within 3 years, FSH >30, and not on hormone therapy. Preclinical CVD was measured using Endo PAT 2000 (Itamar® Medical Ltd) to calculate reactive hyperemic index (RHI) stress to rest pulse amplitude. RHI ≤1.67 indicates endothelial dysfunction. Statistical analysis included analysis of variance for normal distributed variables and Kruskal Wallis test for non-parametric variables. Results The mean age, BMI and estradiol of FHA, no FHA and recent menopause was 26.4 ± 6.2 y, 30.3 ± 3.7 and 53.3 ± 2.6, respectively (p<0.0001); BMI 21.6 ± 6.2 kg/m2, 21.8 ± 2.0, and 23.5 ± 4.2 (p=0.06); and estradiol 29.7±18.2, 46.4 ± 15.7, and 10.9 ± 14.4 pg/ml (p<0.0001), respectively. The median months of FHA amenorrhea was 10.8 months. There were no differences in testosterone (p=0.48), however FHA women had lower leptin (p=0.012) and higher cortisol (p=0.0005) compared to both no FHA and recently menopausal women. Preclinical CVD measured by RHI was significantly lower in FHA compared to no FHA and recently menopausal women (1.8 + 0.5 vs 2.2 + 0.5 vs 2.2 +0.6;p=0.007, respectively). Further 35% of FHA women had RHI ≤1.67 indicating endothelial dysfunction. Conclusion Our results demonstrate endothelial dysfunction in young women with FHA women as compared to eumenorrheic women and older recently menopausal women not on hormone therapy. Our findings suggest that FHA may be an important contributor to CVD mortality in younger women and that preclinical CVD in FHA is not likely due to hypoestrogenemia alone. Future studies will focus on identifying potential treatment targets for CVD prevention. Presentation: Monday, June 13, 2022 12:15 p.m. - 12:30 p.m.
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Alonso, Sylvie, Eve Willery, Genevieve Renauld-Mongénie, and Camille Locht. "Production of Nontypeable Haemophilus influenzae HtrA by Recombinant Bordetella pertussis with the Use of Filamentous Hemagglutinin as a Carrier." Infection and Immunity 73, no. 7 (July 2005): 4295–301. http://dx.doi.org/10.1128/iai.73.7.4295-4301.2005.
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ABSTRACT Bordetella pertussis, the etiologic agent of whooping cough, is a highly infectious human pathogen capable of inducing mucosal and systemic immune responses upon a single intranasal administration. In an attenuated, pertussis toxin (PTX)-deficient recombinant form, it may therefore constitute an efficient bacterial vector that is particularly well adapted for the delivery of heterologous antigens to the respiratory mucosa. Filamentous hemagglutinin (FHA) has been used as a carrier to present foreign antigens at the bacterial surface, thereby inducing local, systemic, and protective immune responses to these antigens in mice. Both full-length and truncated (Fha44) forms of FHA have been used for antigen presentation. To investigate the effect of the carrier (FHA or Fha44) on antibody responses to passenger antigens, we genetically fused the HtrA protein of nontypeable Haemophilus influenzae to either FHA form. The fha-htrA and Fha44 gene-htrA hybrids were expressed as single copies inserted into the chromosome of PTX-deficient B. pertussis. Both chimeras were secreted into the culture supernatants of the recombinant strains and were recognized by anti-FHA and anti-HtrA antibodies. Intranasal infection with the strain producing the FHA-HtrA hybrid led to significantly higher anti-HtrA and anti-FHA antibody titers than those obtained in mice infected with the Fha44-HtrA-producing strain. Interestingly, the B. pertussis strain producing the Fha44-HtrA chimera colonized the mouse lungs more efficiently than the parental, Fha44-producing strain and gave rise to higher anti-FHA antibody titers than those induced by the parental strain.
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Karikari, John A., Ioan Voicu, and Irene Fang. "FHA vs. Subprime Mortgage Originations: Is FHA the Answer to Subprime Lending?" Journal of Real Estate Finance and Economics 43, no. 4 (November 12, 2009): 441–58. http://dx.doi.org/10.1007/s11146-009-9218-7.
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Wagner, Tristan, Gwénaëlle André-Leroux, Valérie Hindie, Nathalie Barilone, María-Natalia Lisa, Sylviane Hoos, Bertrand Raynal, et al. "Structural insights into the functional versatility of an FHA domain protein in mycobacterial signaling." Science Signaling 12, no. 580 (May 7, 2019): eaav9504. http://dx.doi.org/10.1126/scisignal.aav9504.
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Forkhead-associated (FHA) domains are modules that bind to phosphothreonine (pThr) residues in signaling cascades. The FHA-containing mycobacterial protein GarA is a central element of a phosphorylation-dependent signaling pathway that redirects metabolic flux in response to amino acid starvation or cell growth requirements. GarA acts as a phosphorylation-dependent ON/OFF molecular switch. In its nonphosphorylated ON state, the GarA FHA domain engages in phosphorylation-independent interactions with various metabolic enzymes that orchestrate nitrogen flow, such as 2-oxoglutarate decarboxylase (KGD). However, phosphorylation at the GarA N-terminal region by the protein kinase PknB or PknG triggers autoinhibition through the intramolecular association of the N-terminal domain with the FHA domain, thus blocking all downstream interactions. To investigate these different FHA binding modes, we solved the crystal structures of the mycobacterial upstream (phosphorylation-dependent) complex PknB-GarA and the downstream (phosphorylation-independent) complex GarA-KGD. Our results show that the phosphorylated activation loop of PknB serves as a docking site to recruit GarA through canonical FHA-pThr interactions. However, the same GarA FHA–binding pocket targets an allosteric site on nonphosphorylated KGD, where a key element of recognition is a phosphomimetic aspartate. Further enzymatic and mutagenesis studies revealed that GarA acted as a dynamic allosteric inhibitor of KGD by preventing crucial motions in KGD that are necessary for catalysis. Our results provide evidence for physiological phosphomimetics, supporting numerous mutagenesis studies using such approaches, and illustrate how evolution can shape a single FHA-binding pocket to specifically interact with multiple phosphorylated and nonphosphorylated protein partners.
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Pilli, Mehmet, Deniz Seyrek Intas, Ilker Etikan, Pelin Yigitgor, Martin Kramer, Bernd Tellhelm, and Kerstin von Puckler. "The Role of Femoral Head Size and Femoral Head Coverage in Dogs with and without Hip Dysplasia." Veterinary Sciences 10, no. 2 (February 4, 2023): 120. http://dx.doi.org/10.3390/vetsci10020120.
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The subject of hip dysplasia in dogs is still current and preoccupies both animal owners and veterinarians. Major factors affecting the development of the disorder are hip laxity and incongruent joints. Many studies on etiology, pathogenesis, and early diagnosis have been performed to reduce prevalence and select healthy dogs for breeding. The purpose of the present study was to investigate a possible relationship between dysplasia and femoral head area (FHA), femoral coverage by the acetabulum (CFH) and cranio-caudal distance of the dorsal acetabular rim (CrCdAR). Radiographs of a total of 264 skeletally mature dogs with similar physical characteristics (German wirehaired pointers (GWP), German shepherd dogs (GSD) and Labrador retrievers (LAB)) presented for routine hip dysplasia screening were recruited for the study. FHA, CFH and CrCdAR were measured and related to dysplasia status. Evaluations of FHA (p = 0.011), CFH (p < 0.001) and CrCdAR length (p = 0.003) measurements revealed significant interactions between breed, sex and FCI scores, so they had to be assessed separately. The results revealed that FHA tends to decrease as the hip dysplasia score worsens. There was no significant relationship between FHA and dysplasia assessment. FHA is breed-specific and is larger in normal and near-normal male (p = 0.001, p = 0.020) and female (p = 0.001, p = 0.013) GWP compared to GSD, respectively. FHA is greater in normal male GWP (p = 0.011) and GSD (p = 0.040) compared to females. There was a significant and strong positive correlation between FHA and CrCdAR in all breeds and sexes. Additionally, FCI scoring had a medium (GWP, GSD) to strong (LAB) negative correlation with CFH.
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Qu, Hai Bo, Rachel J. Waugh O'Neill, and Mei Wei. "The Effect of Cell Behavior on Apatites with Different Fluorine Contents." Key Engineering Materials 284-286 (April 2005): 577–80. http://dx.doi.org/10.4028/www.scientific.net/kem.284-286.577.
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Fluoridated hydroxyapatite (FHA) discs with various fluorine contents have been used to study the effect of fluorine content on early-stage cell behavior. FHA powders with fluorine content in the range 0-0.577 (mol F /mol apatite) were pressed into discs and sintered at 1200°C for 1 hour. SAOs-2 rat osteosarcoma cells were cultured on each FHA disc and tissue culture polystyrene (control) with the same seeding density for 4 hours. The cell count was conducted using Alamarblue, and the morphology of cell attachment was observed using environmental scanning electron microscopy. It was apparent that the fluorine content in FHA had significant impact on the early cell behavior.
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Gusev, Dmitry V., Sergey Yu Kuznetsov, Tatyana Yu Ivanets, and Galina E. Chernukha. "Differential diagnosis of various forms of functional hypothalamic amenorrhea." Gynecology 21, no. 4 (August 15, 2019): 14–18. http://dx.doi.org/10.26442/20795696.2019.3.190525.
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Aim. To investigate the usefulness of differential diagnostic criteria of functional hypothalamic amenorrhea (FHA) related to energy deficiency and stress. Materials and methods. There were provided clinical and laboratory examination of 56 patients with FHA associated with stressful events (group 1) and 64 patients with FHA on the background of eating disorders (group 2), including assessment of adipose tissue, determination of leptin level, adipose tissue index and differential index. Results. Patients of group 2 were distinguished by a more significant deficiency of body mass index, total body fat, and leptin levels in comparison with patients of group 1. The differential index [AUC=0.907 (0.84-0.97)] turned out to be the most informative indicator in the differential diagnosis of various forms of FHA, its threshold value was 21.4, the least informative - body mass index [AUC=0.78 (0.71-0.87)]. Conclusion. The differential index can be considered as an informative differential diagnostic criterion for various forms of FHA.
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Xie, Changlin, Chao He, Yiyang Jiang, Hailong Yu, Lin Cheng, Gilbert Nshogoza, Moududee Sayed Ala, et al. "Structural insights into the recognition of phosphorylated Hop1 by Mek1." Acta Crystallographica Section D Structural Biology 74, no. 10 (October 1, 2018): 1027–38. http://dx.doi.org/10.1107/s2059798318011993.
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The FHA domain-containing protein Mek1 is a meiosis-specific kinase that is involved in the regulation of interhomolog recombination in meiosis in Saccharomyces cerevisiae. The recruitment and activation of Mek1 require the phosphorylation of the chromosome axis protein Hop1 at Thr318 (pT318), which is necessary for recognition by the Mek1 FHA domain. Here, crystal structures of the Mek1 FHA domain in the apo state and in complex with the Hop1 pT318 peptide are presented, demonstrating that the hydrophobic residues Phe320 and Val321 at the pT+2 and pT+3 positions in the ligand contribute to the preferential recognition. It was further found that in Schizosaccharomyces pombe Mek1 FHA binds both pT15 in its N-terminal SQ/TQ cluster domain (SCD) and pT270 in the Hop1 SCD. The results revealed the structural basis for the preferential recognition of phosphorylated Hop1 by Mek1 in S. cerevisiae and facilitate the understanding of the interaction between the S. pombe Mek1 FHA domain and its binding targets.
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Le Chau, Ngoc, Ngoc Thoai Tran, and Thanh-Phong Dao. "Behavior Analysis of a Flexure Hinge Array." Mathematical Problems in Engineering 2021 (April 13, 2021): 1–11. http://dx.doi.org/10.1155/2021/9947090.
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Compliant mechanisms have been well designed to reach an ultra-high accuracy in positioning systems. However, the displacement of compliant mechanisms is still a major problem that restricts practical applications. Hence, a new flexure hinge array (FHA) is proposed to improve its displacement in this article. This paper is aimed to design and optimize the FHA. The structure of FHA is constructed by series-parallel array. Analytical calculations of the FHA are derived so as to analyze the stiffness and deformation. The displacement of the FHA is optimized by moth-flame optimization algorithm. The results determined that optimal parameters are found at Lt1 of 20.58 mm, w t 1 of 1.92 mm, and w t 2 of 2.29 mm. Besides, the optimal displacement is about 27.02 mm. Through Kruskal–Wallis test, the results verified that the proposed MFO outperforms other optimization algorithms in terms of searching the largest displacement. Validations of the analytical models are verified through simulations and experiments. The theoretical results are close to the experimental results. Additionally, the displacement of the FHA is superior that of existing joints. The displacement in the z-direction is approximately 32 mm according to a displacement of 12 mm in the x-direction.
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Li, Min, Satoshi Komasa, Shigeki Hontsu, Yoshiya Hashimoto, and Joji Okazaki. "Structural Characterization and Osseointegrative Properties of Pulsed Laser-Deposited Fluorinated Hydroxyapatite Films on Nano-Zirconia for Implant Applications." International Journal of Molecular Sciences 23, no. 5 (February 22, 2022): 2416. http://dx.doi.org/10.3390/ijms23052416.
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Standard zirconia implants used in restoration still present problems related to inertness and long-term stability. Various physicochemical approaches have been used to modify the implant surfaces to improve early and late bone-to-implant integration; however, no ideal surface modification has been reported. This study used pulsed laser deposition to deposit a fluorinated hydroxyapatite (FHA) film on a zirconia implant to create a biologically active surface. The film prepared was uniform, dense, and crack-free, and exhibited granular surface droplets; it also presented excellent mechanical strength and favorable biological behavior. The FHA-coated implant was implanted on the femur of Sprague–Dawley rats, and various tests and analyses were performed. Results show that the in vitro initial cell activity on the FHA-coated samples was enhanced. In addition, higher alkaline phosphatase activity and cell mineralization were detected in cells cultured on the FHA-coated groups. Further, the newly formed bone volume of the FHA-coated group was higher than that of the bare micro-adjusted composite nano-zirconia (NANOZR) group. Therefore, the FHA film facilitated osseointegration and may improve the long-term survival rates of dental implants, and could become part of a new treatment technology for implant surfaces, promoting further optimization of NANOZR implant materials.
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Anh Tuyet, Ngo Thi. "CHARACTERIZATION OF FLUORIDATED HYDROXYAPATITE (FHA) SOL-GEL COATINGS ON TITANIUM SUBSTRATE." Vietnam Journal of Science and Technology 55, no. 5B (March 24, 2018): 40. http://dx.doi.org/10.15625/2525-2518/55/5b/12208.
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In this paper, FHA coatings [FHA, Ca10(PO4)6(OH)2-x Fx] (wherein 0 ≤ x ≤ 2) were deposited on titanium substrate by sol-gel method with heat treatment at 900 oC for 4 hours. Different concentrations of F- were incorporated into the apatite structure during the sols preparation. The FHA sols were prepared using various amounts of ammonium fluoride [NH4F] with the [P]/[F] molar ratios of 12, 6, 4, 3 in order to have the corresponding compositions of Ca10(PO4)6(F0.5 OH1.5), Ca10(PO4)6 FOH, Ca10(PO4)6(F1.5 OH0.5) and Ca10(PO4)6F2, respectively. The fabricated FHA coatings were assessed by various methods, namely: morphological structure and chemical composition of coatings were studied by scanning electron microscopy (SEM) and Energy dispersive spectrometry (EDS). The anti-corrosion properties of samples were evaluated by Potentiodynamic polarization curves and Nyquist impedance spectrum. The biocompatibility of FHA coatings on titanium substrates were evaluated by in-vitro tests in simulated body fluid (SBF) solution during 21 days, and ICPMS (Inductively Coupled Plasma Mass Spectrometry) analysis method has been used. The results showed that with dense structure, FHA coatings expressed higher anti-corrosion and biocompatibility performance as compared with that of HA coating.
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Wang, Qianqian, Shize Liu, Xuejun Gao, Yan Wei, Xuliang Deng, Haifeng Chen, and Xuehui Zhang. "Remineralizing Efficacy of Fluorohydroxyapatite Gel on Artificial Dentinal Caries Lesion." Journal of Nanomaterials 2015 (2015): 1–9. http://dx.doi.org/10.1155/2015/380326.
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The aim was to evaluate the remineralizing efficacy of fluorohydroxyapatite (FHA) gel on artificial dentinal caries lesion in vitro. Artificial carious lesions were made on occlusal cavities of teeth by exposing the dentin surface to a demineralizing solution. Each cavity was capped with a 3 mm thick FHA gel for 4 weeks. After the FHA gel was removed, the surface morphology and structure of the dentin were characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR). The dentin mineral density (DMD) was measured by micro-computed tomography (Micro-CT). A layer of dense and orderly hexagonal crystal structure, with average diameter of 1 μm and thickness of 4~5 μm, could be observed on dentin surface. These crystals exhibited elemental peaks for calcium, phosphorus, carbon, and oxygen and characteristic peaks of hydroxyapatite (HA) and fluorapatite (FA) via XRD and FT-IR. The DMD of dentin surface layer significantly increased after it was capped with FHA gel (P<0.05). In the present study, the FHA gel could rapidly construct apatite on the artificial dentin caries surface and significantly increase the mineral density, which suggests that FHA gel might be a proper IPT material with remineralizing function.
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Coutte, Loïc, Sylvie Alonso, Nathalie Reveneau, Eve Willery, Brigitte Quatannens, Camille Locht, and Françoise Jacob-Dubuisson. "Role of Adhesin Release for Mucosal Colonization by a Bacterial Pathogen." Journal of Experimental Medicine 197, no. 6 (March 10, 2003): 735–42. http://dx.doi.org/10.1084/jem.20021153.
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Pathogen attachment is a crucial early step in mucosal infections. This step is mediated by important virulence factors called adhesins. To exert these functions, adhesins are typically surface-exposed, although, surprisingly, some are also released into the extracellular milieu, the relevance of which has previously not been studied. To address the role of adhesin release in pathogenesis, we used Bordetella pertussis as a model, since its major adhesin, filamentous hemagglutinin (FHA), partitions between the bacterial surface and the extracellular milieu. FHA release depends on its maturation by the specific B. pertussis protease SphB1. We constructed SphB1-deficient mutants and found that they were strongly affected in their ability to colonize the mouse respiratory tract, although they adhered even better to host cells in vitro than their wild-type parent strain. The defect in colonization could be overcome by prior nasal instillation of purified FHA or by coinfection with FHA-releasing B. pertussis strains, but not with SphB1-producing FHA-deficient strains, ruling out a nonspecific effect of SphB1. These results indicate that the release of FHA is important for colonization, as it may facilitate the dispersal of bacteria from microcolonies and the binding to new sites in the respiratory tract.
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Muhsinun, Muhsinun, and Mulia Rasyidi. "SINTESIS DAN KARAKTERISASI ASAM LEMAK HIDROKSAMAT DARI EKSTRAK MINYAK MENTAH DEDAK PADI." SAINTEKES: Jurnal Sains, Teknologi Dan Kesehatan 2, no. 1 (January 28, 2023): 74–78. http://dx.doi.org/10.55681/saintekes.v2i1.30.
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Kompleks asam lemak hidroksamat (FHA) dengan beberapa ion logam telah digunakan dalam kimia analitik sebagai reagen untuk gravimetri, spektrofotometri logam, pengkhelat untuk mineral bumi yang langka dan untuk pengekstrak ion-ion logam dari fase air. Oleh sebab itu, sangat perlu dilakukan sintesis FHA dengan bahan dasar yang mengandung asam lemak ini. Salah satu sumber asam lemak adalah minyak mentah dedak padi. Minyak mentah dedak padi mengandung asam lemak dengan rantai sedang dan panjang sehingga memiliki potensi yang sangat besar sebagai bahan baku sintesis asam lemak hidroksamat. Adapun tujuan dari penelitian ini adalah untuk mensintesis FHA dari minyak mentah dedak padi secara enzimatis dan karakterisasi produk FHA yang dihasilkan. Metode yang digunakan adalah metode enzimatis yang meliputi beberapa tahap pengerjaan yaitu tahap sintesis, tahap pemurnian dan tahap karakterisasi. Dari hasil penelitian, diperoleh persentase hasil sintesis asam lemak hidroksamat dari minyak mentah dedak padi setelah dilakukan perbanyakan dengan menggunakan kondisi sintesisnya adalah sekitar 52,37%. Jumlah gugus asam hidroksamat dalam 1 gram sampel kering asam lemak hidroksamat adalah 3,01 mmol. Berdasarkan hasil analisis uji warna dengan CuSO4 dan FeCl3 didapatkan warna kompleks yang khas untuk kedua logam tersebut dengan FHA yaitu warna hijau dan merah tua. Sedangkan dari analisis FTIR, didapatkan spektrum gugus fungsi asam hidroksamat dari sampel FHA.
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Makolle, Sarah, Sophie Catteau-Jonard, Geoffroy Robin, and Didier Dewailly. "Revisiting the serum level of anti-Müllerian hormone in patients with functional hypothalamic anovulation." Human Reproduction 36, no. 4 (February 13, 2021): 1043–51. http://dx.doi.org/10.1093/humrep/deab024.
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Abstract STUDY QUESTION Are serum levels of anti-Müllerian hormone (AMH) normal in patients with functional hypothalamic anovulation (FHA)? SUMMARY ANSWER Our study confirms that in the general FHA population, serum AMH levels are not decreased, but if patients with polycystic ovarian morphology (PCOM) are excluded, levels become significantly lower, as in other situations of gonadotropic insufficiency. WHAT IS KNOWN ALREADY In most situations of low LH (physiological, pharmacological or pathological), serum AMH levels are low. However, paradoxically, many publications have reported normal or even increased serum AMH levels in FHA patients. STUDY DESIGN, SIZE, DURATION Retrospective observational study conducted in an academic centre. The data concerning the study population was collected between 2006 and 2015 from a database including clinical, biological and ultrasound information. PARTICIPANTS/MATERIALS, SETTING, METHODS A total of 45 FHA patients were compared to 37 controls matched based on age and body mass index (BMI). Serum LH, FSH, androstenedione, total testosterone, prolactin and AMH levels were measured by immunoassay. We defined PCOM with strict criteria: a follicle number per ovary (FNPO) ≥ 12 or ≥ 19 per ovary, depending on the date on which the assessment was carried out and the ultrasound device. An AMH level ≥ 35 pmol/l could be a substitute for an excess FNPO. Controls meeting these criteria were not included in this study. MAIN RESULTS AND THE ROLE OF CHANCE There was no significant difference in the ranges of AMH levels between FHA and controls. Using strict criteria to define PCOM status, 46.7% of FHA patients had PCOM. After excluding these patients, the levels of AMH were significantly lower (P < 0.002) in FHA patients compared to controls. Within the FHA group, patients with PCOM had significantly higher ranks of AMH levels and BMI than those without PCOM. However, within the PCOM+ subgroup, the ranks of LH, FSH and A levels were still lower than in controls (P < 0.0001, <0.002 and <0.05, respectively). The positive correlation between AMH and LH was significant in the controls but not in the FHA group. However, in the FHA PCOM+, there was a strong positive correlation between BMI and LH. LIMITATIONS, REASONS FOR CAUTION This is a retrospective study; our controls did not represent the general population as they were recruited in an ART centre; we used a modified classification for PCOM using follicle count and/or AMH level with in-house thresholds to define the follicle excess; the AMH assay used is no longer commercially available. WIDER IMPLICATIONS OF THE FINDINGS Besides biasing the results of AMH assay in FHA patients, the presence of PCOM in FHA patients despite low gonadotropin and androgen levels raises the issue of epigenetically acquired amplification of androgen and/or FSH sensitivity within granulosa cells from polycystic ovaries. In terms of clinical practice, it seems important not to diagnose a low ovarian reserve in FHA patients too quickly on the basis of a decreased AMH level alone. On the contrary, a high AMH level in the context of a menstrual disorder and PCOM should not lead to a misdiagnosis of polycystic ovary syndrome (PCOS) if the basal LH is low. STUDY FUNDING/COMPETING INTEREST(S) None TRIAL REGISTRATION NUMBER N/A
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Franco, A. A., L. Hu, C. J. Grim, G. Gopinath, V. Sathyamoorthy, K. G. Jarvis, C. Lee, et al. "Characterization of Putative Virulence Genes on the Related RepFIB Plasmids Harbored by Cronobacter spp." Applied and Environmental Microbiology 77, no. 10 (March 18, 2011): 3255–67. http://dx.doi.org/10.1128/aem.03023-10.
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ABSTRACTCronobacterspp. are emerging neonatal pathogens that cause meningitis, sepsis, and necrotizing enterocolitis. The genusChronobacterconsists of six species:C. sakazakii,C. malonaticus,C. muytjensii,C. turicensis,C. dublinensis, andCronobactergenomospecies group 1. Whole-genome sequencing ofC. sakazakiiBAA-894 andC. turicensisz3032 revealed that they harbor similarly sized plasmids identified as pESA3 (131 kb) and pCTU1 (138 kb), respectively.In silicoanalysis showed that both plasmids encode a single RepFIB-like origin of replication gene,repA, as well as two iron acquisition systems (eitCBADandiucABCD/iutA). In a chrome azurol S agar diffusion assay, it was demonstrated that siderophore activity was associated with the presence of pESA3 or pCTU1. Additionally, pESA3 contains acpa(Cronobacterplasminogen activator) gene and a 17-kb type 6 secretion system (T6SS) locus, while pCTU1 contains a 27-kb region encoding a filamentous hemagglutinin gene (fhaB), its specifc transporter gene (fhaC), and associated putative adhesins (FHA locus), suggesting that these are virulence plasmids. In arepA-targeted PCR assay, 97% of 229Cronobacterspecies isolates were found to possess a homologous RepFIB plasmid. AllrepAPCR-positive strains were also positive for theeitCBADandiucABCD/iutAiron acquisition systems. However, the presence ofcpa, T6SS, and FHA loci depended on species, demonstrating a strong correlation with the presence of virulence traits, plasmid type, and species. These results support the hypothesis that these plasmids have evolved from a single archetypical plasmid backbone through the cointegration, or deletion, of specific virulence traits in each species.
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Krent, Mollie. "Remediating Racism for Rent: A Landlord’s Obligation Under the FHA." Michigan Law Review, no. 119.8 (2021): 1757. http://dx.doi.org/10.36644/mlr.119.8.remediating.
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The Fair Housing Act (FHA) is an expansive and powerful piece of legislation that furthers equal housing in the United States by ferreting out discrimination in the housing market. While the power of the Act is well recognized by courts, the full contours of the FHA are still to be refined. In particular, it remains unsettled whether and when a landlord can be liable for tenant-on-tenant harassment. This Note argues, first, that the FHA does recognize liability in such a circumstance and, second, that a landlord should be subject to liability for her negligence in such a circumstance. Part I illustrates how the purpose and text of the FHA and analogous civil rights provisions suggest that a landlord should be held liable for her response to tenant-on-tenant harassment. Part II analyzes the standards of liability for tenant-on-tenant harassment that currently exist in the context of the FHA. Part III argues that a negligence standard of liability best accounts for the special status of the home and the unique nature of the landlord-tenant relationship.
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van den Berg, Bernard M., Henry Beekhuizen, Rob J. L. Willems, Frits R. Mooi, and Ralph van Furth. "Role of Bordetella pertussis Virulence Factors in Adherence to Epithelial Cell Lines Derived from the Human Respiratory Tract." Infection and Immunity 67, no. 3 (March 1, 1999): 1056–62. http://dx.doi.org/10.1128/iai.67.3.1056-1062.1999.
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ABSTRACT During colonization of the respiratory tract by Bordetella pertussis, virulence factors contribute to adherence of the bacterium to the respiratory tract epithelium. In the present study, we examined the roles of the virulence factors filamentous hemagglutinin (FHA), fimbriae, pertactin (Prn), and pertussis toxin (PT) in the adherence of B. pertussis to cells of the human bronchial epithelial cell line NCI-H292 and of the laryngeal epithelial cell line HEp-2. Using B. pertussis mutant strains and purified FHA, fimbriae, Prn, and PT, we demonstrated that both fimbriae and FHA are involved in the adhesion of B. pertussis to laryngeal epithelial cells, whereas only FHA is involved in the adherence to bronchial epithelial cells. For PT and Prn, no role as adhesion factor was found. However, purified PT bound to both bronchial and laryngeal cells and as such reduced the adherence of B. pertussis to these cells. These data may imply that fimbriae play a role in infection of only the laryngeal mucosa, while FHA is the major factor in colonization of the entire respiratory tract.
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Nicholson, Tracy L., Susan L. Brockmeier, and Crystal L. Loving. "Contribution of Bordetella bronchiseptica Filamentous Hemagglutinin and Pertactin to Respiratory Disease in Swine." Infection and Immunity 77, no. 5 (February 23, 2009): 2136–46. http://dx.doi.org/10.1128/iai.01379-08.
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ABSTRACT Bordetella bronchiseptica is pervasive in swine populations and plays multiple roles in respiratory disease. Most studies addressing virulence factors of B. bronchiseptica are based on isolates derived from hosts other than pigs. Two well-studied virulence factors implicated in the adhesion process are filamentous hemagglutinin (FHA) and pertactin (PRN). We hypothesized that both FHA and PRN would serve critical roles in the adhesion process and be necessary for colonization of the swine respiratory tract. To investigate the role of FHA and PRN in Bordetella pathogenesis in swine, we constructed mutants containing an in-frame deletion of the FHA or the PRN structural gene in a virulent B. bronchiseptica swine isolate. Both mutants were compared to the wild-type swine isolate for their ability to colonize and cause disease in swine. Colonization of the FHA mutant was lower than that of the wild type at all respiratory tract sites and time points examined and caused limited to no disease. In contrast, the PRN mutant caused similar disease severity relative to the wild type; however, colonization of the PRN mutant was reduced relative to the wild type during early and late infection and induced higher anti-Bordetella antibody titers. Together, our results indicate that despite inducing different pathologies and antibody responses, both FHA and PRN are necessary for optimal colonization of the swine respiratory tract.
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Mielcarek, Nathalie, Inger Nordström, Franco D. Menozzi, Camille Locht, and Jan Holmgren. "Genital Antibody Responses in Mice after Intranasal Infection with an Attenuated Candidate Vector Strain ofBordetella pertussis." Infection and Immunity 68, no. 2 (February 1, 2000): 485–91. http://dx.doi.org/10.1128/iai.68.2.485-491.2000.
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ABSTRACT Intranasal administration of live attenuated Bordetella pertussis, from which the pertussis toxin gene has been deleted, has previously been shown to give rise to high levels of serum immunoglobulin G (IgG) antibodies against both the protective antigen filamentous hemagglutinin (FHA) and heterologous antigens genetically fused to FHA. Here, we extend these results by demonstrating that anti-FHA IgA and IgG antibodies are also produced in the genital tract of mice, both in the vagina and in the uterus, after a single intranasal administration of B. pertussis. By comparing the immune responses induced after infection with wild-type virulentB. pertussis with that induced by infection with an attenuated pertussis toxin-deficient strain, we conclude that pertussis toxin produced by the virulent bacteria does not modify antibody production to FHA in the genital tract of B. pertussis-infected mice. The intranasal infection with either the attenuated or the virulent B. pertussis strain also led to the development of immunologic memory that could be efficiently boosted with purified FHA administered either intranasally or intravaginally to give rise to a significant increase in the levels of specific IgA and IgG produced locally in the genital tract, as well as of specific antibodies in the serum. These observations suggest that attenuatedB. pertussis could be a promising vector for intranasal administration to induce antibody responses against antigens from sexually transmitted pathogens fused to FHA.
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Abramson, Tzvia, Hassya Kedem, and David Relman. "Bordetella pertussis filamentous hemagglutinin induces nuclear translocation of RelB/p52 in U-937 macrophages (135.24)." Journal of Immunology 182, no. 1_Supplement (April 1, 2009): 135.24. http://dx.doi.org/10.4049/jimmunol.182.supp.135.24.
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Abstract Filamentous hemagglutinin (FHA) is a dominant adhesin expressed by the respiratory pathogen Bordetella pertussis that also triggers inflammatory as well as immunosuppressive functions in the host. Given the role of NF-κB transcription factor family members in these functions, we examined the effects of FHA on NF-κB activation at early and late time points in two cell types relevant to natural infection: human bronchial epithelial (BEAS-2B) cells and human macrophage-like (U-937 derived) cells. We recently showed that FHA blocks proteasomal activity in both, the bronchial epithelial cell line and in monocyte-derived macrophages, resulting in accumulation of IκBα at 2 hours or greater incubation times. RelB and alternative NF-κB pathways regulate monopoiesis and monocyte-derived dendritic cell subset development. We report here that incubation of U-937 cells with FHA for more than 4 hours results in reduced levels of cytosolic RelB and increased levels of nuclear RelB and p52, as shown by Western blot analysis. No such effects were observed in bronchial epithelial cells (BEAS-2B). These data suggest that FHA induces nuclear translocation of RelB /p52 and activation of the alternative NF-κB pathway in U-937 cells, but not in BEAS-2B cells, and offer a molecular mechanism by which FHA might affect development and maturation of myeloid host cells.
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Prince, Harry E., Jay M. Lieberman, and James D. Cherry. "Age-Related Differences in Patterns of Increased Bordetella pertussis Antibodies." Clinical and Vaccine Immunology 19, no. 4 (February 22, 2012): 545–50. http://dx.doi.org/10.1128/cvi.05725-11.
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ABSTRACTDuring the period 2008 to 2010, we identified 11,386 serum samples with increased (positive) levels of antibodies recognizingBordetella pertussisantigens. We sought to characterize the distribution of positive antibody result patterns in relation to patient age. IgG and IgA antibodies recognizing pertussis toxin (PT) and filamentous hemagglutinin (FHA) were quantified using a multianalyte immunodetection assay. Four mutually exclusive positive result patterns were observed: increased FHA antibodies only, increased PT IgA but not IgG, increased PT IgG but not IgA, and increased PT IgG and IgA. In patients <21 years old, the predominant pattern was increased PT IgG but not IgA, whereas in patients ≥21 years old, it was increased FHA antibodies only. The proportion of positive serum samples exhibiting increased PT IgA but not IgG was <20% in all age categories but showed a stepwise rise with age. The proportions of positive serum samples exhibiting increased PT IgG and IgA were similar (26 to 32%) in the age categories spanning 11 to 60 years of age but lower in the <11- and >60-year-old groups. In 3 of 5 age categories, a significant rise in the proportion of positive serum samples exhibiting increased FHA antibodies only occurred in 2010. Patterns of positiveB. pertussisantibody results varied with age. The predominance of increased FHA antibodies only in patients >20 years old suggests that many adults thought to haveB. pertussisinfections actually have other infections that induce FHA-reactive antibodies. Similarly, the 2010 rise in the frequency of increased FHA antibodies only in some age groups suggests an increase in non-B. pertussisinfections.
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Bellinzoni, Marco, and Pedro M. Alzari. "Bacterial Metabolism under FHA Control." Structure 17, no. 4 (April 2009): 487–88. http://dx.doi.org/10.1016/j.str.2009.03.004.
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Bolhari, Behnam, Aidin Sooratgar, Maryam Pourhajibagher, Nazanin Chitsaz, and Iman Hamraz. "Evaluation of the Antimicrobial Effect of Mineral Trioxide Aggregate Mixed with Fluorohydroxyapatite against E. faecalis In Vitro." Scientific World Journal 2021 (November 24, 2021): 1–7. http://dx.doi.org/10.1155/2021/6318690.
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Enterococcus faecalis is the dominant microorganism in chronic apical periodontitis. It is more resistant to local antiseptic agents than other endodontic microorganisms. Currently, mineral trioxide aggregate (MTA) is considered as an ideal material in many endodontic procedures. Some studies have shown that MTA has good antibacterial activity against E. faecalis. However, some studies have investigated the effect of incorporating some materials into MTA on its antibacterial activity against E. faecalis. No study has evaluated the effect of incorporating fluorohydroxyapatite nanoparticles (nano-FHA) on the antimicrobial activity of MTA. Therefore, the present study evaluated the antimicrobial effect of MTA mixed with nano-FHA on E. faecalis in vitro. The study was carried out on 18 samples in three groups: pure MTA, MTA mixed with 10 wt% of nano-FHA, and MTA mixed with 15 wt% of nan-FHA. The effect of nano-FHA on the antibacterial activity of MTA on E. faecalis was evaluated by evaluating the growth inhibition zone around each sample. The antimicrobial effect of samples on inhibiting E. faecalis biofilm formation and inhibiting microbial growth of E. faecalis in the planktonic phase was evaluated by disk agar diffusion (DAD), biofilm inhibition assay (BIA), and direct contact assay (DCA) tests, respectively. All the above tests were analyzed after 24 and 72 hours. Factorial designs were used for statistical analyses. Tukey tests were used for two-by-two comparisons. All the statistical analyses were carried out with SPSS 26. DAD results showed no formation of the growth inhibition zone in all the samples after 24 and 72 hours. The microbial colony counts in the BIA and DCA tests in the groups modified with FHA nanoparticles were significantly lower than the pure MTA group ( P < 0.05 ). The microbial colony counts increased in all the groups over time ( P < 0.05 ). Incorporating nano-FHA into MTA improved the antimicrobial activity of MTA against E. faecalis compared to pure MTA. The highest antimicrobial activity was achieved after incorporating 15 wt% of nano-FHA into MTA at the 72-hour interval.
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FATHI, M. H., and E. MOHAMMADI ZAHRANI. "MECHANOCHEMICAL SYNTHESIS OF FLUORIDATED HYDROXYAPATITE NANOPOWDER." International Journal of Modern Physics B 22, no. 18n19 (July 30, 2008): 3099–106. http://dx.doi.org/10.1142/s0217979208047961.
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Recently fluoridated hydroxyapatite (FHA) has been developed since it possesses lower solubility than pure hydroxyapatite (HA), while maintaining the comparable bioactivity and biocompatibility in dental and orthopedic application. The aim of this work was to synthesize and characterize the FHA nanopowder via mechanochemical activation method. Mechanochemical reaction was performed in the planetary ball mill at 300 rpm rotation speed by using 8 balls with 2 cm diameter. XRD technique was used to evaluate phase and composition and determine the grain size of prepared FHA nanopowder. FTIR spectroscopy was utilized to identify the functional groups and to compare obtained powder with bone apatite. The results showed that the synthesis of FHA after 6 hr ball milling at 300 rpm was completed. Fluorhydroxyapatite grain size was almost 37nm after 6 hr of milling.
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Zhu, Qing Xia, Wei Hui Jiang, Hong Da Wang, and Chuan Shao. "Preparing Fluorhydroxyapatite by Aqueous Precipitation Method." Advanced Materials Research 412 (November 2011): 167–70. http://dx.doi.org/10.4028/www.scientific.net/amr.412.167.
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The nanosized fluorhydroxyapatite (FHA) had been synthesized by aqueous precipitation method. The effects of synthesis temperature,fluoride ion concentration and pH value on the fluoride substitution were investigated.The phase composition and the change of crystal structure were characterized by X-ray diffraction and fourier transform infrared spectroscopy. The results show that crystal lattice parameters and bond energy make changes by incorporation of F in the structure.The size of FHA crystals increase as the precipitation temperature. The phase composition of FHA is mainly controlled by the pH value.
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Alonso, Sylvie, Nathalie Reveneau, Kévin Pethe, and Camille Locht. "Eighty-Kilodalton N-Terminal Moiety of Bordetella pertussis Filamentous Hemagglutinin: Adherence, Immunogenicity, and Protective Role." Infection and Immunity 70, no. 8 (August 2002): 4142–47. http://dx.doi.org/10.1128/iai.70.8.4142-4147.2002.
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ABSTRACT Bordetella pertussis, the etiological agent of whooping cough, produces a number of factors, such as toxins and adhesins, that are required for full expression of virulence. Filamentous hemagglutinin (FHA) is the major adhesin of B. pertussis. It is a protein of approximately 220 kDa, found both associated at the bacterial cell surface and secreted into the extracellular milieu. Despite its importance in B. pertussis pathogenesis and its inclusion in most acellular pertussis vaccines, little is known about the functional importance of individual domains in infection and in the induction of protective immunity. In this study, we analyzed the role of the approximately 80-kDa N-terminal domain of FHA, designated Fha44, in B. pertussis adherence, colonization, and immunogenicity. Although Fha44 contains the complete heparan sulfate-binding domain, it is not sufficient for adherence to epithelial cells or macrophages. It also cannot replace FHA during colonization of the mouse respiratory tract. Infection with a B. pertussis strain producing Fha44 instead of FHA does not induce anti-FHA antibodies, whereas such antibodies can readily be induced by intranasal administration of purified Fha44. In addition, mice immunized with purified Fha44 were protected against challenge with wild-type B. pertussis, indicating that Fha44 contains protective epitopes. Compared to FHA, Fha44 is much smaller and much more soluble and is therefore easier to purify and to store. These advantages may perhaps warrant considering Fha44 for inclusion in acellular pertussis vaccines.
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Tondnevis, Farbod, Mohammadali Ketabi, Reza Fekrazad, Ali Sadeghi, and Mohamad Mahdi Abolhasani. "Using Chitosan Besides Nano Hydroxyapatite and Fluorohydroxyapatite Boost Dental Pulp Stem Cell Proliferation." Journal of Biomimetics, Biomaterials and Biomedical Engineering 42 (July 2019): 39–50. http://dx.doi.org/10.4028/www.scientific.net/jbbbe.42.39.
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The dental tissue scaffold must provide a favorable surface for dental pulp stem cell attachment and proliferation. Employing nanohydroxyapatite (HA) and nanofluorohydroxyapatite (FHA) beside synthetic and organic polymer in favor of scaffolds would be used in bone and dental tissue engineering. In this research, nanoHA and FHA/chitosan scaffolds were synthesized by freeze-drying technique. Surface morphology, chemical composition and hydrophilicity have a great impact on initial cell attachment which will further affect the cell viability and proliferation which evaluated by SEM, XRD and contact angle measurement. Bioactivity of scaffolds was investigated by immersion in simulated body fluid (SBF) and cell proliferation assay. In freeze-drying technique percentage usage of hydroxyapatite could be risen up to 40% and shown better macro-mechanical and physical properties and bioactivity. According to obtained results by adding chitosan, contact angle was decreased by %54 and %37 for polycaprolactone (PCL)/HA and PCL/FHA scaffolds. In addition, addition of chitosan causes significant increase in the cell proliferation for PCL/HA and PCL/FHA up to 81% and 164%, respectively. These results indicate that PCL/FHA/chitosan scaffold represent a big potential for dental tissue engineering.
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Robin, G., C. Gallo, S. Catteau-Jonard, C. Lefebvre-Maunoury, P. Pigny, A. Duhamel, and D. Dewailly. "Polycystic Ovary-Like Abnormalities (PCO-L) in Women with Functional Hypothalamic Amenorrhea." Journal of Clinical Endocrinology & Metabolism 97, no. 11 (November 1, 2012): 4236–43. http://dx.doi.org/10.1210/jc.2012-1836.
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Context: In the general population, about 30% of asymptomatic women have polycystic ovary-like abnormalities (PCO-L), i.e. polycystic ovarian morphology (PCOM) at ultrasound and/or increased anti-Müllerian hormone (AMH) serum level. PCOM has also been reported in 30–50% of women with functional hypothalamic amenorrhea (FHA). Objective: The aim of this study was to verify whether both PCOM and excessive AMH level indicate PCO-L in FHA and to elucidate its significance. Design: We conducted a retrospective analysis using a database and comparison with a control population. Setting: Subjects received ambulatory care in an academic hospital. Patients: Fifty-eight patients with FHA were compared to 217 control women with nonendocrine infertility and body mass index of less than 25 kg/m2. Interventions: There were no interventions. Main Outcome Measures: We measured serum testosterone, androstenedione, FSH, LH, AMH, and ovarian area values. The antral follicle count (AFC) was used as a binary variable (i.e. negative or positive) because of the evolution of its sensitivity over the time of this study. The ability of these variables (except AFC) to detect PCO-L in both populations was tested by cluster analysis. Results: One cluster (cluster 2) suggesting PCO-L was detected in the control population (n = 52; 24%), whereas two such clusters were observed in the FHA population (n = 22 and n = 6; 38 and 10%; clusters 2 and 3, respectively). Cluster 2 in FHA had similar features of PCO-L as cluster 2 in controls, with higher prevalence of positive AFC (70%) and PCOM (70%), higher values of ovarian area and higher serum AMH (P < 0.0001 for all), and testosterone levels (P < 0.01) than in cluster 1. Cluster 3 in FHA was peculiar, with frankly elevated AMH levels. In the whole population (controls + FHA), PCO-L was significantly associated with lower FSH values (P < 0.0001). Conclusion: PCO-L in FHA is a frequent and usually incidental finding of unclear significance, as in controls. The association of PCO-L with hypothalamic amenorrhea should not lead to a mistaken diagnosis of PCOS.
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Wang, Bin, Shifan Yang, Lei Zhang, and Zheng-Guo He. "Archaeal Eukaryote-Like Serine/Threonine Protein Kinase Interacts with and Phosphorylates a Forkhead-Associated-Domain-Containing Protein." Journal of Bacteriology 192, no. 7 (January 29, 2010): 1956–64. http://dx.doi.org/10.1128/jb.01471-09.
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ABSTRACT Protein phosphorylation plays an important role in cell signaling. However, in the Archaea, little is known about which proteins are phosphorylated and which kinases are involved. In this study, we identified, for the first time, a typical eukaryote-like Ser/Thr protein kinase and its protein partner, a forkhead-associated (FHA)-domain-containing protein, from the archaeon Sulfolobus tokodaii strain 7. This protein kinase, ST1565, physically interacted with the FHA-domain-containing protein, ST0829, both in vivo and in vitro. ST1565 preferred Mn2+ as a cofactor for autophosphorylation and for substrate phosphorylation; the optimal temperature for this was 45°C, and the optimal pH was 5.5 to 7.5. The critical amino acid residues of the conserved FHA and kinase domain sites were identified by performing a series of mutation assays. Thr329 was part of a major activation site in the kinase, while Thr326 was a negative regulation site. Several mutants with amino acid substitutions in the conserved FHA domain sites of ST0829 did not physically interact with ST1565. A structural model for the FHA domain demonstrated that the mutation sites were located at the edge of the protein and thus were in the domain that potentially interacts with ST1565. This report describes pioneering work on the third domain of life, the Archaea, showing that a protein kinase interacts with and phosphorylates an FHA-domain-containing protein. Our data provide critical information on the structural or functional characteristics of archaeal proteins and could help increase our understanding of fundamental signaling mechanisms in all three domains of life.
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Millo, Christian, Carlo Bravo, Stefano Covelli, Elena Pavoni, Elisa Petranich, Marco Contin, Maria De Nobili, et al. "Metal Binding and Sources of Humic Substances in Recent Sediments from the Cananéia-Iguape Estuarine-Lagoon Complex (South-Eastern Brazil)." Applied Sciences 11, no. 18 (September 12, 2021): 8466. http://dx.doi.org/10.3390/app11188466.
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The Cananéia-Iguape estuarine–lagoon complex (São Paulo state, Brazil) is a natural laboratory to study metal binding by humic substances (HS) in subtropical settings. This transitional environment is evolving into a freshwater environment due to water input from the Ribeira River, funneled through the Valo Grande Canal (Iguape). Past mining activities in the Ribeira River basin and maritime traffic are suspected to be potential sources of trace metals in the system. In this study, the trace metal contents of Free Humic Acids (FHA), Bound Humic Acids (BHA), and Fulvic Acids (FA) extracted from sedimentary organic matter were investigated. Moreover, the sources of HS were traced using their stable carbon isotope compositions and C/N ratios. The results suggested a mixed marine–terrestrial source of FHA, BHA, and FA. Copper and Cr were the most abundant trace metals bound to HS. On average, Cu showed concentrations of 176, 115, and 37.9 μg g−1 in FHA, BHA, and FA, respectively, whereas Cr showed average concentrations of 47.4, 86.3, and 43.9 μg g−1 in FHA, BHA, and FA, respectively. Marine FHA showed the highest binding capacity for trace metals, whereas terrestrial FA derived from the decay of mangrove organic matter showed the lowest binding capacity.
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de Lange, A., R. Huiskes, and J. M. G. Kauer. "Effects of Data Smoothing on the Reconstruction of Helical Axis Parameters in Human Joint Kinematics." Journal of Biomechanical Engineering 112, no. 2 (May 1, 1990): 107–13. http://dx.doi.org/10.1115/1.2891160.
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In biomechanical joint-motion analyses, the continuous motion to be studied is often approximated by a sequence of finite displacements, and the Finite Helical Axis (FHA) or “screw axis” for each displacement is estimated from position measurements on a number of anatomical or artificial landmarks. When FHA parameters are directly determined from raw (noisy) displacement data, both the position and the direction of the FHA are ill-determined, in particular when the sequential displacement steps are small. This implies, that under certain conditions, the continuous pathways of joint motions cannot be adequately described. The purpose of the present experimental study is to investigate the applicability of smoothing (or filtering) techniques, in those cases where FHA parameters are ill-determined. Two different quintic-spline smoothing methods were used to analyze the motion data obtained with Roentgenstereophotogrammetry in two experiments. One concerning carpal motions in a wrist-joint specimen, and one relative to a kinematic laboratory model, in which the axis positions are a priori known. The smoothed and nonsmoothed FHA parameter errors were compared. The influences of the number of samples and the size of the sampling interval (displacement step) were investigated, as were the effects of equidistant and nonequidistant sampling conditions and noise invariance.
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Bashkirov, Vladimir I., Elena V. Bashkirova, Edwin Haghnazari, and Wolf-Dietrich Heyer. "Direct Kinase-to-Kinase Signaling Mediated by the FHA Phosphoprotein Recognition Domain of the Dun1 DNA Damage Checkpoint Kinase." Molecular and Cellular Biology 23, no. 4 (February 15, 2003): 1441–52. http://dx.doi.org/10.1128/mcb.23.4.1441-1452.2003.
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ABSTRACT The serine-threonine kinase Dun1 contains a forkhead-associated (FHA) domain and functions in the DNA damage checkpoint pathway of Saccharomyces cerevisiae. It belongs to the Chk2 family of checkpoint kinases, which includes S. cerevisiae Rad53 and Mek1, Schizosaccharomyces pombe Cds1, and human Chk2. Dun1 is required for DNA damage-induced transcription of certain target genes, transient G2/M arrest after DNA damage, and DNA damage-induced phosphorylation of the DNA repair protein Rad55. Here we report that the FHA phosphoprotein recognition domain of Dun1 is required for direct phosphorylation of Dun1 by Rad53 kinase in vitro and in vivo. trans phosphorylation by Rad53 does not require the Dun1 kinase activity and is likely to involve only a transient interaction between the two kinases. The checkpoint functions of Dun1 kinase in DNA damage-induced transcription, G2/M cell cycle arrest, and Rad55 phosphorylation are severely compromised in an FHA domain mutant of Dun1. As a consequence, the Dun1 FHA domain mutant displays enhanced sensitivity to genotoxic stress induced by UV, methyl methanesulfonate, and the replication inhibitor hydroxyurea. We show that the Dun1 FHA domain is critical for direct kinase-to-kinase signaling from Rad53 to Dun1 in the DNA damage checkpoint pathway.
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Hussein, Ahmad H., Elisabeth M. Davis, Scott A. Halperin, and Song F. Lee. "Construction and Characterization of Single-Chain Variable Fragment Antibodies Directed against the Bordetella pertussis Surface Adhesins Filamentous Hemagglutinin and Pertactin." Infection and Immunity 75, no. 11 (August 27, 2007): 5476–82. http://dx.doi.org/10.1128/iai.00494-07.
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ABSTRACT A single-chain variable fragment (scFv) antibody library against Bordetella pertussis was constructed using M13 phage display. The library was enriched for phages surface displaying functional scFv by biopanning against B. pertussis immobilized on polystyrene plates. Two hundred eighty-eight individual clones from the enriched library were screened for binding to B. pertussis cells, filamentous hemagglutinin (FHA), and pertactin (PRN) in enzyme-linked immunosorbent assays (ELISAs). Based on the binding ability, the clones were put into eight groups. The scFv DNA inserts from the 288 clones were digested with BstOI, and 18 unique restriction patterns, named types 1 to 18, were found. Eight clones (types 1 to 7 and 18) were selected for further testing against FHA, PRN, and B. pertussis by ELISA. The results showed that types 1, 5, 7, and 18 bound strongly to B. pertussis cells as well as FHA and PRN. Type 3 bound strongly to the cells and FHA but weakly to PRN. Types 4 and 6 bound FHA only, and type 2 did not bind to the cells or antigens. The ability of the eight clones to inhibit B. pertussis from binding to HEp-2 cells was assayed. Types 1, 5, and 7, but not the remaining clones, inhibited the adherence of B. pertussis to HEp-2 cells. The scFvs were sequenced, and the deduced amino acid sequence showed that the scFvs were different antibodies. Maltose-binding protein (MBP) fusion proteins composed of three different regions of FHA (heparin-binding domain, carbohydrate recognition domain, and the RGD triplet motif) were constructed. The three fusion proteins and Mal85 (MBP-FHA type I domain) were used to map the binding sites for scFvs of types 1, 5, and 7 by ELISA. The results showed that all three scFvs bound to the heparin-binding domain fusion protein but not the other fusion proteins. BALB/c mice who received recombinant phage-treated B. pertussis had reduced bacterial counts in the nasal cavity, trachea, and lungs compared to the control groups.