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Biomedical subjects

J Bruno

Publications and source records attributed to J Bruno.

At least 19 recordsLinked to original sources
PubMed2006

Magnetic resonance signal, rather than tendon volume, correlates to pain and functional impairment in chronic Achilles tendinopathy.

PURPOSE: To depict abnormal tendon matrix composition using magnetic resonance imaging (MRI) in chronic Achilles tendinopathy, and correlate intratendinous signal alterations to pain and functional impairment. MATERIAL AND METHODS: MRI of the Achilles tendon was performed on 25 patients with chronic Achilles tendinopathy (median age 50, range 37-71 years). All patients suffered from pain in the mid-portion of the Achilles tendon. Intratendinous signal was calculated from five different sagittal sequences, using a computerized 3D seed-growing technique. Pain and functional impairment were evaluated using a questionnaire completed by patients. RESULTS: Severity of pain and functional impairment correlated to increased mean intratendinous signal in the painful tendon in all MR sequences (P < 0.05, median r = 0.38, range 0.28-0.43 for pain; P < 0.05, median r = 0.48, range 0.29-0.49 for functional impairment). However, tendon volume did not correlate to pain or functional impairment (P > 0.05). Difference in mean intratendinous signal between symptomatic and contralateral asymptomatic tendons was highly significant in all sequences (P < 0.05) except on T2-weighted images (P = 0.6). CONCLUSION: Severity of pain and disability correlated to increased MR signal rather than to tendon volume in patients with unilateral mid-portion chronic Achilles tendinopathy.

Achilles Tendon↗
PubMed2006

Dynamic evolution of antibody populations in a rhesus macaque infected with attenuated simian immunodeficiency virus identified by surface plasmon resonance.

BACKGROUND: Increasing evidence suggests that an effective AIDS vaccine will need to elicit broadly neutralizing antibody responses. However, the mechanisms of antibody-mediated neutralization have not been defined. Previous studies from our lab have identified significant differences in the rates of antibody binding to trimeric SIV envelope proteins that correlate with neutralization sensitivity. Importantly, these results demonstrate differences in monoclonal antibody (MAb) binding to neutralization-sensitive and neutralization-resistant envelope proteins, suggesting that one mechanism for virus neutralization may be related to the stability of antibody binding. To date, little has been done to evaluate the binding properties of polyclonal serum antibodies elicited by SIV infection or vaccination. METHODS: In the current study, we translate these findings with MAbs to study antibody binding properties of polyclonal serum antibody responses generated in rhesus macaques infected with attenuated SIV. Quantitative and qualitative binding properties of well-characterized longitudinal serum samples to trimeric, recombinant SIV gp140 envelope proteins were analyzed using surface plasmon resonance (SPR) technology (Biacore). RESULTS: Results from these studies identified two antibody populations in most of the samples analyzed; one antibody population exhibited fast association/dissociation rates (unstable) while the other population demonstrated slower association/dissociation rates (stable). Over time, the percentage of the total binding response of each antibody population evolved, demonstrating a dynamic evolution of the antibody response that was consistent with the maturation of antibody responses defined using our standard panel of serological assays. However, the current studies provided a higher resolution analysis of polyclonal antibody binding properties, particularly with respect to the early time-points post-infection (PI), that is not possible with standard serological assays. More importantly, the increased stability of the antibody population with time PI corresponded with potent neutralization of homologous SIV in vitro. CONCLUSIONS: These results suggest that the stability of the antibody-envelope interaction may be an important mechanism of serum antibody virus neutralization. In addition, measurements of the 'apparent' rates of association and dissociation may offer unique numerical descriptors to characterize the level of antibody maturation achieved by candidate vaccine strategies capable of eliciting broadly neutralizing antibody responses.

Amino Acid Sequence↗
PubMed2005

The use of a high-FeO olivine rock as a redox buffer in a nuclear waste repository.

Due to the higher stability of the spent nuclear fuel (mainly composed of UO2) under reducing conditions, and in order to enhance the retention/retardation of some key radionuclides, the olivine rock from the Lovasjärvi intrusion has been proposed as a potential redox-active backfill-additive in deep high-level nuclear waste (HLNW) repositories. In this work, two different approaches have been undertaken in order to establish the redox buffer capacity of olivine rock: (1) The capacity of the rock to respond to changes in pH or pe has been demonstrated and the final (pH, pe) coordinates agree with the control exerted by the system Fe(II)/Fe(III). (2) The rate of consumption of oxygen has been determined at different pH values. These rates are higher than the ones reported in the literature for other solids, what would point to the possibility of using this rock as an additive to the backfill material in a HLNW.

Buffers↗
PubMed2000

Purification, characterization and molecular cloning of prophenoloxidases from Sarcophaga bullata.

Prophenoloxidase (PPO) is a key enzyme associated with both melanin biosynthesis and sclerotization in insects. This enzyme is involved in three physiologically important processes viz., cuticular hardening, defense reactions and wound healing in insects. It was isolated from the larval hemolymph of Sarcophaga bullata and purified by employing ammonium sulfate precipitation, Phenyl Sepharose chromatography, DEAE-Sepharose chromatography, and Sephacryl S-200 column chromatography. The purified enzyme exhibited two closely moving bands on 7.5% SDS-PAGE under denaturing conditions. From the estimates of molecular weight on Sephacryl S-100, TSK-3000 HPLC column and SDS-PAGE, which ranged from 90,000 to 100,000, it was inferred that the enzyme is made up of a single polypeptide chain. Activation of PPO (K(a)=40 microM) was achieved by the cationic detergent, cetyl pyridinium chloride below its critical micellar concentration (0.8 mM) indicating that the detergent molecules are binding specifically to the PPO and causing the activation. Neither anionic, nor nonionic (or zwitterionic) detergents activated the PPO. The active enzyme exhibited wide substrate specificity and marked thermal unstability. Using primers designed to conserved amino acid sequences from known PPOs, we PCR amplified and cloned two PPO genes from the sarcophagid larvae. The clones encoded polypeptides of 685 and 691 amino acids. They contained two distinct copper binding regions and lacked the signal peptide sequence. They showed a high degree of homology to dipteran PPOs. Both contained putative thiol ester site, two proteolytic activation sites and a conserved C-terminal region common to all known PPOs.

Amino Acid Sequence↗
PubMed1998

Nitrate reductase alters 3-nitrotyrosine accumulation and cell cycle progression in LPS + IFN-gamma-stimulated RAW 264.7 cells.

Nitrite (NO2-), an end product of nitrogen radical metabolism, has recently been shown to increase tyrosine nitration by activated leukocytes indicating that nitrite modulates the immune response. We investigated the hypothesis that nitrite may increase nitration of molecular targets within activated cells leading to altered cell cycle progression. Intracellular nitrite was increased by transfection of murine macrophage-like RAW 264.7 cells with the nitrate reductase gene obtained from barley. Nitrate reductase facilitates the conversion of nitrate to nitrite; thus when extracellular nitrate is present, intracellular nitrite will be increased. Results show that addition of KNO3 increases NO2- production and intracellular nitrotyrosine accumulation in the transfectant but not the parent. Inhibition of nitric oxide synthesis with L-NAME during activation with IFN-gamma + LPS reduced NO2- production to the same extent in both cell lines; however, cellular accumulation of nitrotyrosine was reduced by only 25% in the transfectant (P = 0.21) and 49% in the parent cell line (P = 0.007), suggesting that intracellular nitrite increased nitrotyrosine accumulation through a pathway not requiring NO synthesis, i.e., myeloperoxidase system. Approximately 15% of the transfected cells had 4n DNA content 24 h postactivation compared to < 1% of the parent cells. Increased DNA copy number was correlated to nitrotyrosine accumulation. These findings show that intracellular nitrite can increase accumulation of nitrotyrosine and that nitration is linked to cell cycle perturbation.

Animals↗
PubMed1998

Scintigraphic evaluation of malignant melanoma lesions with Tc-99m tetrofosmin.

Two cases of malignant melanoma (primary and metastatic lesions) imaged with Tc-99m tetrofosmin are reported. One patient showed intense uptake in a primary skin lesion of the thorax, and the other patient had accumulation in skin, cerebellum, breast, and lymph node metastases. Like Tc-99m MIBI, Tc-99m tetrofosmin imaging may be of clinical relevance in the evaluation of suspicious skin lesions and in patients with known cutaneous malignant melanoma in the assessment of recurrent disease during follow-up evaluation.

Adult↗
PubMed1998

Clinical validation of ineffective breathing pattern, ineffective airway clearance, and impaired gas exchange.

PURPOSE: To describe the clinical validation of symptoms or defining characteristics of three respiratory diagnoses. The contributing factors or etiologies of the diagnoses were identified and the degree of importance of 30 nursing interventions, 15 direct care and 15 teaching, was rated for each diagnosis and each patient. Three nursing diagnoses--ineffective breathing pattern (IBP), ineffective airway clearance (IAC), and impaired gas exchange (IGE)--were among the most frequently used, yet no reported clinical studies validated the defining characteristics of these diagnoses. This study answers the research questions: What are the defining characteristics of IBP, IAC, and IGE? What are the etiologies of IBP, IAC, AND IGE? What are the most important interventions for IBP, IAC, and IGE? DESIGN: Standardized clinical validation using a convenience sample of 76 people hospitalized with medical and surgical diagnoses, in one U.S. city, and identified as having one of the three diagnoses. Data were collected in 1992-1993. METHODS: A literature-based concept analysis generated 37 possible defining characteristics for the three diagnoses which were included in the instrument. The nurse experts conducted a health history and physical examination of each patients and decided (a) whether the 37 defining characteristics were present or absent, (b) the degree of importance of each possible defining characteristic for making one or more of the diagnoses, (c) the etiologies, and (d) which of the 30 nursing interventions were important for each diagnosis and patient. FINDINGS: For each diagnosis, many of the 37 possible defining characteristics were judged as present but few reached the criterion of .50 as important for making one of the diagnoses. Two of the possible defining characteristics reached this criterion for IBP, seven for IAC, and two for IGE. In contrast to the defining characteristics approved by NANDA, the subjective cues of "expresses fatigue" and "expresses anxiety" were judged as important for making one or more of the diagnoses. CONCLUSIONS: Clinical validation methods allow discriminating among defining characteristics. Data that are present are not necessarily characteristic of a diagnosis, and the subjective cues of expresses fatigue or anxiety may be important for making these diagnoses.

Adult↗
PubMed1998

Myeloma light chains are ligands for cubilin (gp280).

Although myeloma light chains are known to undergo receptor-mediated endocytosis in the kidney, the molecular identity of the receptor has not been characterized. We examined the interaction between cubilin (gp280) and four species of light chains isolated from the urine of patients with multiple myeloma. Four lines of evidence identify cubilin, a giant glycoprotein receptor, which is restricted in distribution to endocytic scavenger pathways and which has potent effects on endosomal trafficking, as a potentially physiologically relevant binding site for light chains: 1) light chains coeluted during immunoaffinity purification of cubilin; 2) polyclonal antisera to cubilin but not control sera, displaced human light chain binding from rat renal brush-border membranes; 3) cubilin bound to multiple species of light chains during surface plasmon resonance; 4) anti-cubilin antiserum interfered with light chain endocytosis by visceral yolk sac epithelial cells. However, both binding of light chains to brush-border membranes and endocytosis of light chains by yolk sac epithelial cells were only partially inhibited by anticubilin antibodies, suggesting presence of additional or alternate binding sites for light chains. Excess light chain had a potent inhibitory effect on endosomal fusion in vitro. Binding showed dose and time-dependent saturability with low-affinity, high-capacity equilibrium binding parameters. These data demonstrate that cubilin plays a role in the endocytosis and trafficking of light chains in renal proximal tubule cells.

Amino Acid Sequence↗
PubMed1997

A major cis activator of the IRBP gene contains CRX-binding and Ret-1/PCE-I elements.

PURPOSE: Interphotoreceptor retinoid binding protein (IRBP) is expressed exclusively and to high levels in photoreceptive cells. This study was an attempt to delineate the minimal regulated control region of the murine IRBP promoter involved in this expression pattern. METHODS: Fragments of the mouse IRBP 5' flanking region were tested for promoter activity in transient transfections of embryonic chick retina cells in primary culture. Electrophoretic mobility shift assays were used to identify specific cis-acting DNA elements within these fragments. RESULTS: Nested deletion analysis of a 1783 bp fragment of the murine IRBP 5' flanking region shows that high promoter activity is maintained with truncated fragments as short as 70 bp 5' to transcription start, but is lost with truncation to 45 bases. The 1783 bp promoter is active in cultures of retina cells but not brain cells or fibroblasts. The 70 bp fragment is active in retina and brain cells but not fibroblasts. Within retina cell cultures, the 1783 bp fragment is active in photoreceptor-like and amorphous or unidentifiable cells whereas the 70 bp is additionally active in multipolar neuron-like cells. The -70 to -45 interval contains Ret-1/PCE-I (AATTAG in the IRBP gene), a proposed retina-specific consensus sequence cis element, and a same-strand reversed copy of this sequence, GATTAA, the consensus binding element of the photoreceptor-specific trans-acting factor CRX. Mutation of either element suppresses promoter activity. Paralleling promoter tissue-specificity, the -70 to -45 fragment binds a sequence-specific protein complex found in retina and brain extracts but not fibroblasts. Mutation of both or either element inhibits this binding. CONCLUSIONS: These data suggest that a trans-acting complex binds a cis-element in the -70 to -45 sequence. This binding fully activates transcription but confers only partial tissue-specificity to IRBP gene expression.

Animals↗
PubMed1997

Anionic- and lipophilic-mediated surface binding inhibitors of human leukocyte elastase.

We report the synthesis of a series of diphenylmethane-based oligomers containing anionic and lipophilic functionalities that are potent inhibitors of human leukocyte elastase (HLE). The enzyme inhibition is regulated by the size of the oligomer, as well as, the number of charges. Lipophilicity is an important element in determining potency and specificity against other basic enzymes. Compounds whose scaffolds contain three phenoxyacetic acid groups and three alkyl ethers are competitive and specific inhibitors of HLE with Ki = 20 nM. The mechanism of action of this class of compounds is believed to involve multidendate interactions with the surface of HLE near the active site which prevents substrate access to the catalytic site.

Benzhydryl Compounds↗
PubMed1997

The 5' flanking regions of IRBP and arrestin have promoter activity in primary embryonic chicken retina cell cultures.

Primary cultures of embryonic chicken cells from various tissues were transiently transfected with plasmid vectors containing reporter genes linked to a 1.8 kb fragment of the mouse interphotoreceptor retinoid-binding protein (IRBP) 5' flanking region, a 1.5 kb fragment of the mouse arrestin 5' flanking region, or a 3.4 kb sequence of the bovine arrestin 5' flanking region. Promoter activity was evident in retina-derived cells, but not in fibroblasts or cells from whole brain. Transfection response also varied with transfection method, plasmid DNA concentration, post-transfection incubation time, and cell density. The data suggest that the primary embryonic chicken retinal cell culture system is a useful tool in studying photoreceptor-specific gene regulation.

Animals↗
PubMed1997

Recurrent synovial chondromatosis treated with meniscectomy and synovectomy.

Synovial chondromatosis is a rare benign intraarticular metaplasia of synovium. This process may result in the production of detached particles of highly cellular cartilage in the involved joint spaces. It is most often reported in the larger joints of the body including the knee, hip, elbow, and ankle. Since Axhausen in 1993 reported the first case affecting the temporomandibular joint, several articles have been listed in the literature regarding the presentation, diagnosis, and management of this form of an arthropathy. This is a case of a recurrent synovial chondromatosis that was approached with a meniscectomy and a complete synovectomy.

Arthroscopy↗
PubMed1996

Characterization of Eu(III) binding to a series of calmodulin binding site mutants using laser-induced Eu(III) luminescence spectroscopy.

Laser-induced luminescence techniques were used in a rigorous evaluation of the Eu(3+)-binding behavior of a recombinant (Drosophila melanogaster) calmodulin and a series of calmodulin binding site mutants in which the bidentate glutamic acid residue in position 12 of each metal ion binding loop is systematically replaced with lysine. For the range of Ca2+ concentrations at which calmodulin functions (10(-5)-10(-6) M), Ca2+ binding is effectively eliminated at the mutated site; however, the luminescence studies show that the Eu3+ ion binds to the modified site with reduced affinity. The mutations do not significantly change the intermetal ion distances from their wild type values. These were determined by Eu3+-->Nd3+ Förster-type non-radiative energy transfer experiments. Consistent with the results of Ca(2+)-binding studies, mutation of sites II and IV in the N- and C-terminal domains, respectively, produces a larger alteration in the Eu(3+)-luminescence and Eu(3+)-binding behavior than does mutation of sites I and III. Modification of either of the sites in the C-terminus (III or IV, numbered from the amino terminus) causes two additional H2O molecules (four H2O molecules total) to bind to the Eu3+ ion in order to compensate for the loss of the bidentate glutamic acid residue. Consequently, the partner site in the domain loses an H2O molecule, thereby coordinating another ligand from the protein. Mutation of either of the high-affinity Ln(3+)-binding sites (I or II) has global effects on the Eu(3+)-binding behavior of the protein molecule.

Animals↗
PubMed1996

Formulation and antitumor activity evaluation of nanocrystalline suspensions of poorly soluble anticancer drugs.

PURPOSE: Determine if wet milling technology could be used to formulate water insoluble antitumor agents as stabilized nanocrystalline drug suspensions that retain biological effectiveness following intravenous injection. METHODS: The versatility of the approach is demonstrated by evaluation of four poorly water soluble chemotherapeutic agents that exhibit diverse chemistries and mechanisms of action. The compounds selected were: piposulfan (alkylating agent), etoposide (topoisomerase II inhibitor), camptothecin (topoisomerase I inhibitor) and paclitaxel (antimitotic agent). The agents were wet milled as a 2% w/v solids suspension containing 1% w/v surfactant stabilizer using a low energy ball mill. The size, physical stability and efficacy of the nanocrystalline suspensions were evaluated. RESULTS: The data show the feasibility of formulating poorly water soluble anticancer agents as physically stable aqueous nanocrystalline suspensions. The suspensions are physically stable and efficacious following intravenous injection. CONCLUSIONS: Wet milling technology is a feasible approach for formulating poorly water soluble chemotherapeutic agents that may offer a number of advantages over a more classical approach.

Animals↗
PubMed1995

Prognostic value of histological grading in ductal adenocarcinoma of the pancreas. Klöppel vs TNM grading.

A new histological grading system with prognostic correlation for pancreatic cancer was proposed by Klöppel et al. in 1985. Histological sections from 60 ductal adenocarcinomas operated on between January 1980 and December 1990 were retrospectively reviewed in order to compare Klöppel's grading with standard TNM's grading and assess their prognostic value. Klöppel grading was determined through the following histologic and cytologic factors: number duct-like structures, mucus production, neoplastic epithelium, arrangement and pleomorphism of nuclei, and mitotic activity. A score from 0 (well differentiated) to 2 (poorly differentiated) was given to each factor. The mean value obtained dividing the sum of the different values by the number of parameters was used to construct a malignancy scale and therefore allocate each patient to his Klöppel grading. The concordance index K between the two grading systems was relevant (K = 0.85 p < 0.001). There was no relation either between gradings (Klöppel or TNM) and preoperative duration of symptoms or between gradings and UICC stages. TNM's G2 grades of malignancy, N status, and tumor stage were significantly related to survival time (p < 0.05). Klöppel's grading does not show any advantage over the classical and simpler TNM's grading, even though it can be considered more objective and therefore more easily reproducible. This characteristic further should be enhanced by the introduction of a malignancy scale such as the "mean value."

Aged↗