Solid-phase development of a L-hydroxybenzotriazole linker for heterocycle synthesis using analytical constructs.

The development of a 1-hydroxybenzotriazole linker for the synthesis of heterocyclic derivatives is described, utilizing analytical construct methodology to facilitate the analysis of resin samples. A UV-chromophore-containing analytical construct enabled the accurate determination of resin loading and the automated monitoring of key reactions using only small quantities of resin. The syntheses of an array of isoxazole derivatives are reported.

Primary oral health care in black Americans: an assessment of current status and future needs.

To improve health for all in the United States by the year 2000, dental health needs must be considered a component of total health and primary care. The failure to address dental needs has reached a crisis level, particularly in the black and underserved communities throughout the nation. Data from several nationwide studies have shown that oral disease is greater in black Americans than their white counterparts. More severe periodontal disease patterns, untreated dental decay, and earlier tooth loss were observed. Key minority subgroups received less preventive care.

Characterization of second-generation monoclonal antibodies against carcinoembryonic antigen.

BACKGROUND: A second-generation panel of anti-carcinoembryonic antigen (CEA) monoclonal antibodies (MoAb) has been generated, and the specificity has been compared with that of the first panel of MoAb used to differentiate meconium antigen (MA) from CEA. METHODS AND RESULTS: Four of the MoAb had similar specificities to the first-generation panel of NP MoAb. MN-15, like its first-generation equivalent, NP-1, reacts with normal cross-reactive antigen (NCA), MA, and CEA; both MN-15 and NP-1 react strongly with granulocytes. MN-2 has properties similar to Class II NP-2, being reactive with MA and CEA, cross-blocking binding to CEA by NP-1, and having low reactivity with granulocytes; both NP-2 and MN-2 stain granulocytes in frozen tissue sections but show minimal staining of granulocytes in sections fixed in formaldehyde solution and embedded in Paraplast (Fischer Scientific, Pittsburgh, PA). MN-14 demonstrates properties similar to the Class III anti-CEA-specific MoAb, NP-4, being unreactive with NCA and MA. MN-14, as compared with NP-4, demonstrated significantly superior tumor targeting in a human colon tumor xenograft model and consistently stronger staining of frozen sections of colon cancer. A fifth MoAb, MN-3, had properties uniquely different from the NP series of MoAb, reacting strongly with granulocytes but not demonstrating the liquid-phase ion-sensitivity binding of CEA exhibited by MN-15 and NP-1. CONCLUSIONS: MN-14 is being evaluated for radioimmunodetection of and radioimmunotherapy for CEA-containing cancers, whereas MN-3 is being studied for the radioimmunodetection of occult infections and sites of inflammation.

Identification of initiating agents in myoglobin-induced lipid peroxidation.

Reaction of metmyoglobin with peroxides is known to involve the formation, possibly via a porphyrin radical-cation, of a ferryl (iron(IV)-oxo) species and a protein radical; there is a little information available as to which of these species initiates the damage which is observed on incubating membrane fractions with such mixtures. It is shown in this study that the initial protein radical, which is a tyrosine phenoxyl radical centered at position 103 in the protein, does not react rapidly with erythrocyte membranes. However a tyrosine peroxyl radical, formed by addition of oxygen to this species, does react rapidly, and it is concluded that this radical, together with the ferryl species, is an important initiating species in myoglobin-induced lipid peroxidation.

The formation of free radicals by cardiac myocytes under oxidative stress and the effects of electron-donating drugs.

The interaction of myoglobin with H2O2 leads via a two-electron oxidation process to the formation of ferryl myoglobin. Metmyoglobin is more readily activated than oxymyoglobin to the ferryl states, which are capable of inducing peroxidative damage to membranes. E.p.r. and optical spectroscopic studies show that the thiol-containing compounds N-(2-mercaptopropionyl) glycine and N-acetylcysteine and the trihydroxamate desferrioxamine attenuate these processes by reducing the ferryl myoglobin species to metmyoglobin, with the formation of thiyl radicals and the desferrioxamine nitroxide radical respectively. Biochemical investigations of the potential for myoglobin in ruptured myocytes to be involved in radical generation, when under oxidative stress, and of the nature of the resulting species, were also undertaken. E.p.r. spectroscopic studies revealed the formation of a radical species which is capable of inducing membrane lipid peroxidation. The interaction of the thiol compounds and desferrioxamine with components of myocardial tissue under these conditions results in the generation of thiol-derived radical species and the desferrioxamine nitroxide radical respectively. These data, along with those obtained using optical spectrocopy, support the assignment of the identity of the radical species generated from the myocytes as the ferryl myoglobin radical.

Preclinical evaluation of an "instant" 99mTc-labeling kit for antibody imaging.

This communication presents information on the quality and stability of a new conjugate of 99mTc to an anticancer antibody fragment and results of animal imaging and distribution studies. Clinical studies have proved that this labeled antibody can image tumors within 2 hours of i.v. injection. It is produced using an "instant kit," merely requiring addition of [99mTc]pertechnetate to a lyophilized vial containing the fragment and a pertechnetate-reducing agent. Within 5 min of addition of pertechnetate to the vial containing the fragment, [99mTc]pertechnetate is reduced and quantitatively bound to an intrinsic high-affinity receptor present in the Fab'. The Fab'-bound 99mTc is completely stable to challenge with diethylenetriaminepentaacetic acid and human serum albumin. Minimal oxidation and release of 99mTc bound to the fragment were observed when the labeled fragment was incubated for 24 h at 37 degrees C. Animal biodistribution studies were consistent with the known metabolic elimination of a radiolabeled Fab' fragment by the kidney. Tumor localization studies in athymic nude mice bearing xenografts of human colonic carcinoma (GW-39) predicted the positive results later demonstrated in clinical studies.

Comparison of tumor targeting in nude mice by murine monoclonal antibodies directed against different human colorectal cancer antigens.

Tumor targeting of five radioiodinated murine monoclonal antibodies (MAbs) directed against human colorectal cancer were studied in nude mice bearing the GW-39 human colonic tumor xenograft. All of the MAbs are of the IgG1 isotype. Two of the MAbs (NP-4 and MN-14) are directed against a Class III carcinoembryonic antigen-specific epitope, but they differ 10-fold in their affinity. The other three MAbs recognize mucins found in colonic cancer. Mu-9 recognizes a peptide determinant similar to that described previously for a polyclonal goat anti-colon-specific antigen p. G9 identifies an organ-specific, tumor-associated carbohydrate epitope. The tumor targeting of these MAbs was compared to that of B72.3, another anti-mucin MAb. The tumor uptake of all the MAbs were similar on days 1, 3, and 7, with an average maximum accretion of between 30 and 40%/g tumor occurring by day 3. This tumor uptake was maintained for 14 days with the anti-mucin MAbs, whereas the percentage of injected dose/g in the tumor for 2 anti-carcinoembryonic antigen MAbs decreased 2-fold by day 14. Although no statistical difference could be found between the percentage of injected dose/g in the tumor for NP-4 and MN-14 (carcinoembryonic antigen MAbs), in a paired-labeled study using 131I-MN-14 and 125I-NP-4, MN-14 uptake in the tumor was consistently 1.3 times higher than that of NP-4 on all days tested. F(ab')2 fragments showed lower tumor uptake (maximum uptake for NP-4 and Mu-9 was 11% on day 1), but the faster clearance resulted in a 4- to 40-fold increase in tumor/blood ratios on day 3 in comparison to the whole IgGs. Fab' fragments had the lowest tumor uptake of the 3 forms of immunoglobulin, with a maximum of only 5%/g 6 h after injection. However, tumor/blood ratios on day 1 for the Fab' fragments were improved 3-fold over that of F(ab')2. All of these MAbs, except Mu-9, identify epitopes that can be detected in plasma, but none of the MAbs complexed appreciably when mixed in vitro with plasma containing antigen at antigen/MAb ratios anticipated to be encountered most frequently in imaging or therapy applications in humans. However, complexation of the MAbs will occur if antigen in the plasma is markedly elevated. These studies suggest that if used clinically, tumor targeting of colorectal cancer with any of these MAb IgG may be similar if these antigens are present at relatively similar concentrations in tumor.(ABSTRACT TRUNCATED AT 400 WORDS)

Murine monoclonal antibody adsorbed onto vinylidene fluoride floccules used to eliminate antibody interference in "sandwich"-type immunoassays.

We have previously reported that human anti-mouse IgG antibody (HAMA) can cause false-positive and false-negative results in "sandwich"-type monoclonal antibody (MAb) assays. To eliminate HAMA interference in "sandwich"-type MAb assays, we investigated the use of MAb on solid-phase, vinylidene fluoride floccules, which we have previously used as a solid-phase second antibody for RIA. The simple procedure effectively removes greater than 95% of HAMA from the most positive serum we have obtained from patients hyperimmunized to murine MAb, and it allows for accurate quantification of carcinoembryonic antigen. The solid-phase complex, added to blood, effectively removes HAMA and (or) "HAMA-type" heterophilic antibody from the sera or plasma.

Solving the problem of antibody interference in commercial "sandwich"-type immunoassays of carcinoembryonic antigen.

We evaluated the effect of human anti-murine antibodies (HAMA) on apparent concentrations of carcinoembryonic antigen (CEA) as measured in serum with commercial enzyme immunoassay (EIA) kits manufactured by Abbot ("two-step" double monoclonal antibody assay), Roche, and Hybritech (room-temperature protocol). In sera from patients given parenteral murine monoclonal antibody for experimental diagnostic and immunotherapy studies, HAMA titers were determined with Immunomedics' "ImmuSTRIP HAMA-EIA" kit reagents. "True" CEA titers were established by using the ImmuCEA/MA-EIA and heat-extraction to destroy HAMA before assay for CEA. The concordance of the ImmuCEA/MA assay with the Abbott and Roche CEA EIAs was established with sera from normal individuals and from patients who had not received parenteral injections of murine monoclonal antibody. At high (100 mg/L) concentrations of HAMA, false-positive results were observed with all three kits. The Hybritech and Roche assays were more sensitive to interference by HAMA than was the Abbott CEA-EIA, false-positive results being observed at HAMA concentrations between 1 and 10 mg/L. Similar sensitivity of the three kits to interference by primate anti-MAb sera was demonstrated. Use of primate anti-MAb sera to create controls with HAMA activity and of analyte is recommended to evaluate MAb assays for potential HAMA interference and for use to devise methods to eliminate HAMA interference.

Factors in the admission decision: skilled nursing facilities in New York State.

Little is known about the policies, practices, or procedures that affect the selection of applicants for nursing home admission. Results of a study of skilled nursing facilities (SNFs) in the state of New York revealed that fewer than 5% of the respondents viewed their goal as merely keeping patients comfortable; about half of the facilities reported their goal as preventing further decline. More than 70% of the respondents stated that skilled nursing beds were in short supply in the area served by their facility, while 91% of the respondents reported that their geographic area had a backlog of patients in acute care hospitals awaiting placement in SNFs. The authors discuss the study results in terms of the respondents' type of facility--public, proprietary, or voluntary. They also discuss admissions personnel, multidisciplinary teams, information collection, family involvement, and other factors that influence admission (eg, philosophy of care). The authors conclude that further research is needed to help coordinate the medical and social needs of patients and their families with the administrative needs of the facility.

Community acceptance of the mentally ill in foster family care.

Community acceptance has been described as essential to the successful placement of the mentally ill in the community. The study reported here examined acceptance of mental patients living in family care homes and found that residents were well accepted by neighbors and by others in the community, albeit on a superficial level, and that acceptance could be predicted by variables relating to the residents' social network.

Admission procedures in skilled nursing facilities.

Little is known about the policies, practices, or procedures that affect the placement of applicants for nursing home care. Toseland and Newman report how financial resources and various forms of physical, mental, and behavioral problems affects the chances that older persons will be admitted to or excluded from nursing facilities.

Classification decisions within psychogeriatric day care.

Classification decisions play a central role in implementation of day care services. A case study is presented which examines classification decisions within an outpatient multilevel psychogeriatric day care program at a state psychiatric center. Staff cited as relevant criteria: social isolation, severity of psychopathology, home emotional climate, activity, ADL and mental status. Issues related to utilization of criteria, such as relevance, consensus, definition and program changes, are discussed.

Evaluation of baboon antiserum to carcinoembryonic antigen.

The common baboon (Papio), immunized with carcinoembryonic antigen (CEA), was found to form specific-site antibodies for CEA. The baboon antiserum does not recognize colon carcinoma antigen-III, but does demonstrate similar ionic strength effects to those previously described for goat antisera to the CEA-specific site(s). Baboon specific-site antibody for CEA can be used for radioimmunoassay of CEA in plasma and gives similar results to the commercial (Roche) goat anti-CEA.

Foster-family care for the elderly: surrogate family or mini-institution?

The rationale behind adult foster care (AFC) has been that "participation in the life of the family" is superior to institutionalization. The extent to which AFC is familial has been widely debated, ranging from claims that this environment provides a surrogate family, to claims that it is no more than a mini-institution. This paper discusses the extent to which elderly clients are integrated into the family and the method by which such integration was measured in a sample of one hundred adult foster homes in New York State. The four dimensions used to measure familism were Affection, Social Interaction, the performance of Ritual, and the minimization of Social Distance.