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1.
Animals (Basel) ; 13(4)2023 Feb 17.
Article in English | MEDLINE | ID: mdl-36830515

ABSTRACT

The aim of this study was to assess if molasses could modify VFA production and the rumen microbial community in vitro. Three beet (treatment Beet) and three cane (treatment Cane) molasses preparations were randomly selected from a variety of samples collected worldwide and incubated in vitro with rumen fluid along with a control sample (treatment CTR, in which no molasses was used). Flasks for VFA analysis were sampled at 0, 1, 2, 3, 4, 6, 8, and 24 h of each incubation. For microbiota analysis, samples from each fermentation flask after 12 and 24 h were subjected to microbial DNA extraction and V3-V4 16S rRNA gene sequencing on an Illumina MiSeq platform. Total net VFA production was higher in the beet and cane preparations than in the control (CTR) group at 24 h (33 mmol/L, 34 mmol/L, and 24.8 mmol/L, respectively), and the composition of VFAs was affected by the inclusion of molasses: acetic acid increased in the CTR group (73.5 mol%), while propionic acid increased in the beet and cane molasses (19.6 mol% and 18.6 mol%, respectively), and butyric acid increased, especially in the cane group (23.2 mol%). Molasses even influenced the composition of the rumen microbiota, and particularly the relative abundance of the most dominant family in the rumen, Prevotellaceae, which decreased compared to CTR (37.13%, 28.88%, and 49.6%, respectively). In contrast, Streptococcaceae (19.62% and 28.10% in molasses compared to 6.23% in CTR), Veillonellaceae (6.48% and 8.67% in molasses compared to 4.54% in CTR), and Fibrobacteraceae (0.90% and 0.88% in molasses compared to 0.62% in CTR) increased in the beet and cane groups compared to the CTR group. Another important finding is the lower proportion of Methanobacteriaceae following the addition of molasses compared to CTR (0.26%, 0.28%, and 0.43%, respectively). This study showed the impact of molasses in influencing VFA production and composition as a result of a modified rumen microbial composition.

3.
PLoS One ; 17(3): e0264458, 2022.
Article in English | MEDLINE | ID: mdl-35294466

ABSTRACT

As people, animals and materials are transported across increasingly large distances in a globalized world, threats to our biosecurity and food security are rising. Aotearoa New Zealand is an island nation with many endemic species, a strong local agricultural industry, and a need to protect these from pest threats, as well as the economy from fraudulent commodities. Mitigation of such threats is much more effective if their origins and pathways for entry are understood. We propose that this may be addressed in Aotearoa using strontium isotope analysis of both pests and products. Bioavailable radiogenic isotopes of strontium are ubiquitous markers of provenance that are increasingly used to trace the origin of animals and plants as well as products, but currently a baseline map across Aotearoa is lacking, preventing use of this technique. Here, we have improved an existing methodology to develop a regional bioavailable strontium isoscape using the best available geospatial datasets for Aotearoa. The isoscape explains 53% of the variation (R2 = 0.53 and RMSE = 0.00098) across the region, for which the primary drivers are the underlying geology, soil pH, and aerosol deposition (dust and sea salt). We tested the potential of this model to determine the origin of cow milk produced across Aotearoa. Predictions for cow milk (n = 33) highlighted all potential origin locations that share similar 87Sr/86Sr values, with the closest predictions averaging 7.05 km away from their true place of origin. These results demonstrate that this bioavailable strontium isoscape is effective for tracing locally produced agricultural products in Aotearoa. Accordingly, it could be used to certify the origin of Aotearoa's products, while also helping to determine if new pest detections were of locally breeding populations or not, or to raise awareness of imported illegal agricultural products.


Subject(s)
Strontium Isotopes , Strontium , Animals , Biosecurity , Humans , New Zealand , Strontium/analysis , Strontium Isotopes/analysis
4.
J Dairy Sci ; 103(7): 6244-6249, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32331893

ABSTRACT

Beet and cane molasses are produced worldwide as a by-product of sugar extraction and are widely used in animal nutrition. Due to their composition, they are fed to ruminants as an energy source. However, molasses has not been properly characterized in the literature; its description has been limited to the type (sugarcane or beet) or to the amount of dry matter (DM), total or water-soluble sugars, crude protein, and ash. Our objective was to better characterize the composition of cane and beet molasses, examine possible differences, and obtain a proper definition of such feeds. For this purpose, 16 cane and 16 beet molasses samples were sourced worldwide and analyzed for chemical composition. The chemical analysis used in this trial characterized 97.4 and 98.3% of the compounds in the DM of cane and beet molasses, respectively. Cane molasses contained less DM compared with beet molasses (76.8 ± 1.02 vs. 78.3 ± 1.61%) as well as crude protein content (6.7 ± 1.8 vs. 13.5 ± 1.4% of DM), with a minimum value of 2.2% of DM in cane molasses and a maximum of 15.6% of DM in beet molasses. The amount of sucrose differed between beet and cane molasses (60.9 ± 4.4 vs. 48.8 ± 6.4% of DM), but variability was high even within cane molasses (39.2-67.3% of DM) and beet molasses. Glucose and fructose were detected in cane molasses (5.3 ± 2.7 and 8.1 ± 2.8% of DM, respectively), showing high variability. Organic acid composition differed as well. Lactic acid was more concentrated in cane molasses than in beet molasses (6.1 ± 2.8 vs. 4.5 ± 1.8% of DM), varying from 1.6 to 12.8% of DM in cane molasses. Dietary cation-anion difference showed numerical differences among cane and beet molasses (7 ± 53 vs. 66 ± 45 mEq/100 g of DM, on average). It varied from -76 to +155 mEq/100 g of DM in the cane group and from +0 to +162 mEq/100 g of DM in the beet group. Data obtained in this study detailed differences in composition between sources of molasses and suggested that a more complete characterization could improve the use of molasses in ration formulation.


Subject(s)
Beta vulgaris/chemistry , Molasses/analysis , Saccharum/chemistry
6.
Vaccine ; 20(3-4): 545-53, 2001 Nov 12.
Article in English | MEDLINE | ID: mdl-11672921

ABSTRACT

In a double-blinded, randomized trial, human immunodeficiency virus (HIV)-infected adults with > or = 200 CD4 cells/microl received placebo (PL), 7-valent conjugate, or 23-valent pneumococcal polysaccharide (PS) vaccine in one of the following two-dose combinations given 8 weeks apart: conjugate-conjugate, conjugate-polysaccharide, placebo-polysaccharide, placebo-placebo. A total of 67 persons completed the study. Neither significant increases in HIV viral load nor severe adverse reactions occurred in any group. After controlling for confounders, when compared with persons receiving placebo-polysaccharide, persons receiving conjugate-conjugate and conjugate-polysaccharide had higher antibody concentrations (serotypes 4, 6B, 9V and serotype 23F, respectively) and opsonophagocytic titers (functional antibody assay, serotypes 9V, 23F and serotypes 4, 6B, 9V, respectively) after the second dose (P<0.05). The second dose with either conjugate or polysaccharide following the first conjugate dose, however, produced no further increase in immune responses.


Subject(s)
Antibodies, Bacterial/blood , HIV Infections/immunology , Pneumococcal Vaccines/immunology , Adult , CD4 Lymphocyte Count , Enzyme-Linked Immunosorbent Assay , HIV Infections/virology , Humans , Phagocytosis , Pneumococcal Vaccines/adverse effects , Vaccines, Conjugate/immunology , Viral Load
7.
J Clin Microbiol ; 38(6): 2043-50, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10834951

ABSTRACT

Pneumococcal conjugate vaccines will eventually be licensed after favorable results from phase III efficacy trials. After licensure of a conjugate vaccine for invasive pneumococcal disease in infants, new conjugate vaccines will likely be licensed primarily on the basis of immunogenicity data rather than clinical efficacy. Analytical methods must therefore be developed, evaluated, and validated to compare immunogenicity results accurately within and between laboratories for different vaccines. At present no analytical technique is uniformly accepted and used in vaccine evaluation studies to determine the acceptable level of agreement between a laboratory result and the assigned value for a given serum sample. This multicenter study describes the magnitude of agreement among 12 laboratories quantifying an identical series of 48 pneumococcal serum specimens from 24 individuals (quality-control sera) by a consensus immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) developed for this study. After provisional or trial antibody concentrations were assigned to the quality-control serum samples for this study, four methods for comparison of a series of laboratory-determined values with the assigned concentrations were evaluated. The percent error between assigned values and laboratory-determined concentrations proved to be the most informative of the four methods. We present guidelines that a laboratory may follow to analyze a series of quality-control sera to determine if it can reproduce the assigned antibody concentrations within an acceptable level of tolerance. While this study focused on a pneumococcal IgG ELISA, the methods that we describe are easily generalizable to other immunological assays.


Subject(s)
Antibodies, Bacterial/blood , Bacterial Vaccines/immunology , Enzyme-Linked Immunosorbent Assay/statistics & numerical data , Streptococcus pneumoniae/immunology , Bacterial Capsules/immunology , Confidence Intervals , Enzyme-Linked Immunosorbent Assay/methods , Evaluation Studies as Topic , Guidelines as Topic , Humans , Models, Statistical , Pneumococcal Infections/prevention & control , Quality Control , Streptococcus pneumoniae/classification , Vaccination
8.
Nurs Sci Q ; 13(1): 60-3, 2000 Jan.
Article in English | MEDLINE | ID: mdl-11847783

ABSTRACT

This article describes leadership and scholarship from the Neuman systems model as requisite for a true practice model necessary for 21st-century scientific professional nursing. A rationale for the integration of leadership and scholarship is provided along with markers to guide the leadership-scholarship protocol, which can be viewed as a scholarly research process. An example from a community nursing center demonstrating the application of the integration of leadership-scholarship is presented to support the proposed integration.


Subject(s)
Community Health Nursing/organization & administration , Continuity of Patient Care/organization & administration , Health Promotion/organization & administration , Holistic Health , Leadership , Models, Nursing , Nurse's Role , Nursing Research/organization & administration , Nursing Theory , Systems Theory , Aged , Community Health Nursing/education , Forecasting , Humans , Job Description , Knowledge , Professional Autonomy , Professional Competence/standards
9.
JAMA ; 281(3): 243-8, 1999 Jan 20.
Article in English | MEDLINE | ID: mdl-9918479

ABSTRACT

CONTEXT: Revaccination of healthy adults with pneumococcal polysaccharide vaccine (PPV) within several years of first vaccination has been associated with a higher than expected frequency and severity of local injection site reactions. The risk of adverse events associated with revaccination of elderly and chronically ill persons 5 or more years after first vaccination, as is currently recommended, has not been well defined. OBJECTIVE: To determine whether revaccination with PPV at least 5 years after first vaccination is associated with more frequent or more serious adverse events than those following first vaccination. DESIGN: Comparative intervention study conducted between April 1996 and August 1997. PARTICIPANTS: Persons aged 50 to 74 years either who had never been vaccinated with PPV (n = 901) or who had been vaccinated once at least 5 years prior to enrollment (n = 513). INTERVENTION: PPV vaccination. MAIN OUTCOME MEASURES: Postvaccination local injection site reactions and prevaccination concentrations of type-specific antibodies. RESULTS: Those who were revaccinated were more likely than those who received their first vaccinations to report a local injection site reaction of at least 10.2 cm (4 in) in diameter within 2 days of vaccination: 11% (55/513) vs 3% (29/901) (relative risk [RR], 3.3; 95% confidence interval [CI], 2.1-5.1). These reactions resolved by a median of 3 days following vaccination. The highest rate was among revaccinated patients who were immunocompetent and did not have chronic illness: 15% (33/228) compared with 3% (10/337) among comparable patients receiving their first vaccinations (RR, 4.9; 95% CI, 2.4-9.7). The risk of these local reactions was significantly correlated with prevaccination geometric mean antibody concentrations. CONCLUSIONS: Physicians and patients should be aware that self-limited local injection site reactions occur more frequently following revaccination compared with first vaccination; however, this risk does not represent a contraindication to revaccination with PPV for recommended groups.


Subject(s)
Bacterial Vaccines/administration & dosage , Bacterial Vaccines/adverse effects , Streptococcus pneumoniae/immunology , Aged , Antibodies, Bacterial/biosynthesis , Bacterial Vaccines/immunology , Female , Humans , Immunization Schedule , Immunocompetence , Immunocompromised Host , Logistic Models , Male , Middle Aged , Multivariate Analysis , Pneumococcal Vaccines , Time Factors
10.
N Engl J Med ; 338(26): 1861-8, 1998 Jun 25.
Article in English | MEDLINE | ID: mdl-9637804

ABSTRACT

BACKGROUND: Outbreaks of pneumococcal disease are uncommon and have occurred mainly in institutional settings. Epidemic, invasive, drug-resistant pneumococcal disease has not been seen among adults in the United States. In February 1996, there was an outbreak of multidrug-resistant pneumococcal pneumonia among the residents of a nursing home in rural Oklahoma. METHODS: We obtained nasopharyngeal swabs for culture from residents and employees. Streptococcus pneumoniae isolates were serotyped and compared by pulsed-field gel electrophoresis. A retrospective cohort study was conducted to identify factors associated with colonization and disease. RESULTS: Pneumonia developed in 11 of 84 residents (13 percent), 3 of whom died. Multidrug-resistant S. pneumoniae, serotype 23F, was isolated from blood and sputum from 7 of the 11 residents with pneumonia (64 percent) and from nasopharygeal specimens from 17 of the 74 residents tested (23 percent) and 2 of the 69 employees tested (3 percent). All the serotype 23F isolates were identical according to pulsed-field gel electrophoresis. Recent use of antibiotics was associated with both colonization (relative risk, 2.3; 95 percent confidence interval, 1.3 to 4.2) and disease (relative risk, 3.6; 95 percent confidence interval, 1.2 to 10.8). Only three residents (4 percent) had undergone pneumococcal vaccination. After residents received pneumococcal vaccine and prophylactic antibiotics, there were no additional cases of pneumonia, and the rates of carriage decreased substantially. CONCLUSIONS: In this outbreak a single pneumococcal strain was disseminated among the residents and employees of a nursing home. The high prevalence of colonization with a virulent organism in an unvaccinated population contributed to the high attack rate. Clusters of pneumococcal disease may be underrecognized in nursing homes, and wider use of pneumococcal vaccine is important to prevent institutional outbreaks of drug-resistant S. pneumoniae infection.


Subject(s)
Bacteremia/epidemiology , Disease Outbreaks , Drug Resistance, Multiple , Nursing Homes , Pneumococcal Infections/epidemiology , Pneumonia, Pneumococcal/epidemiology , Streptococcus pneumoniae/isolation & purification , Adult , Aged , Aged, 80 and over , Bacteremia/microbiology , Bacterial Vaccines , Cohort Studies , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Nasopharynx/microbiology , Pneumococcal Infections/microbiology , Pneumonia, Pneumococcal/microbiology , Retrospective Studies , Risk Factors , Streptococcus pneumoniae/classification , Streptococcus pneumoniae/drug effects
11.
Cancer Pract ; 5(4): 213-9, 1997.
Article in English | MEDLINE | ID: mdl-9250077

ABSTRACT

PURPOSE: This study was conducted to determine the frequency of breast self-examination (BSE), clinical breast examination, and mammography of adult daughters of women with breast cancer. Additionally, the relationships among frequency of self-examination, clinical examination, perceived risk, fear of breast cancer, and frequency of talking with their mothers about breast cancer were assessed. METHODS: A retrospective, correlational descriptive design was used. Questionnaires were mailed to members of a breast cancer support group and to women diagnosed with breast cancer in one medical oncology practice. These women were asked to mail the questionnaires to their adult daughters. RESULTS: There was a significant relationship between frequency of BSE and frequency of talking with mothers about breast cancer. Frequency of self-examination was related inversely to fear of breast cancer. Fear of breast cancer appears to act as a barrier to action whereas frequency of talking with their mothers about breast cancer seems to act as a cue to action in support of the Health Belief Model. CLINICAL IMPLICATIONS: Healthcare providers should make every effort to optimize the practice of BSE in daughters of women with breast cancer. Only 52% reported performing BSE monthly, with the remaining 48% performing BSE less frequently or not at all. Thirty-one percent reported having no formal or printed instruction regarding BSE. Health professionals caring for women who have a family history of breast cancer should assess the educational needs of these women and provide opportunities for them to acquire and demonstrate skills. Periodic re-evaluation of BSE is needed to reinforce importance and demonstrate technique. The development of educational materials developed specifically for daughters of women with breast cancer may be useful in diminishing the perception of an unrealistically high risk of developing breast cancer. With the decrease in fear, which appears to be acting as a barrier to BSE in this group, better breast cancer detection practices in daughters may be realized. Counseling about realistic risk of developing breast cancer also may be useful in reducing the amount of fear of breast cancer in these women. This is an unnecessary burden for any woman to bear and may interfere with her optimal practice of breast cancer detection practices.


Subject(s)
Breast Neoplasms/prevention & control , Breast Self-Examination , Health Knowledge, Attitudes, Practice , Mammography , Nuclear Family/psychology , Adult , Breast Neoplasms/genetics , Female , Humans , Middle Aged , Retrospective Studies , Surveys and Questionnaires
12.
J Infect Dis ; 175(1): 84-90, 1997 Jan.
Article in English | MEDLINE | ID: mdl-8985200

ABSTRACT

Lack of primary immune response in severe combined immunodeficient (SCID) mice engrafted with human peripheral blood lymphocytes (hu-PBL) has limited the applicability of this model. Use of human cytokines, in particular interleukin (IL)-12, was studied in the hu-PBL-SCID model. SCID mice were treated with IL-12 and reconstituted with hu-PBL in T replacement factor. The hu-PBL-SCID mice were immunized with serogroup C meningococcal polysaccharide (MCPS). The MCPS-specific antibody response was determined by ELISA. Thirteen of the 15 immunized, IL-12-treated hu-PBL-SCID mice demonstrated a primary human antibody response to MCPS ranging from 0.25 to 3.3 microg/mL, while no MCPS-specific antibody response was detectable in the 18 controls. Expression of cross-reactive idiotypic markers found on human anti-MCPS antibodies in the immunized hu-PBL-SCID mice was similar to that observed in immunized volunteers.


Subject(s)
Antibodies, Bacterial/biosynthesis , Antigens, T-Independent/immunology , B-Lymphocytes/immunology , Interleukin-12/pharmacology , Lymphocyte Transfusion , Neisseria meningitidis/immunology , Polysaccharides, Bacterial/immunology , Severe Combined Immunodeficiency/immunology , Adult , Animals , Antibodies, Anti-Idiotypic/immunology , Antibodies, Bacterial/blood , B-Lymphocytes/transplantation , Enzyme-Linked Immunosorbent Assay , Hemocyanins/immunology , Humans , Immunization , Kinetics , Mice , Mice, Inbred ICR , Mice, SCID , Transplantation, Heterologous
13.
Clin Diagn Lab Immunol ; 3(1): 84-8, 1996 Jan.
Article in English | MEDLINE | ID: mdl-8770509

ABSTRACT

An interlaboratory study was conducted to determine whether an enzyme-linked immunosorbent assay (ELISA) with an antigen preparation composed of various-sized fragments of Haemophilus influenzae type b polysaccharide conjugated to human serum albumin could be standardized across laboratories and whether the ELISA-derived results from different laboratories are equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-H, influenzae type b polysaccharide antibodies. Twenty coded human serum samples were quantitated by ELISA in 11 laboratories and by RABA in 5 laboratories. The mean RABA-derived values served as the basis for all comparisons. While the overall correspondence of antibody values between the two methods was good, significant differences were found among some of the 11 ELISA data sets and among the mean RABA values. Seven laboratories generated higher ELISA antibody values for low-titered sera. Four laboratories generated antibody concentrations that were not statistically different between the two assay methods. The results therefore indicate that the ELISA can tolerate substantial variations in protocol, such as the use of different plates and different antibody reagents, without affecting the quantitation of serum antibodies. However, attention should be focused on low-titered sera, as some assay conditions may yield spurious results. This ELISA is a serologic assay which can serve as an alternative to the RABA for quantitation of antibodies to H. influenzae type h polysaccharide.


Subject(s)
Antibodies, Bacterial/analysis , Enzyme-Linked Immunosorbent Assay/methods , Haemophilus Vaccines/immunology , Haemophilus influenzae/immunology , Polysaccharides, Bacterial/immunology , Adult , Antibodies, Bacterial/blood , Antigens, Bacterial , Bacterial Capsules , Child , Enzyme-Linked Immunosorbent Assay/statistics & numerical data , Evaluation Studies as Topic , Humans , Laboratories , Radioimmunoassay/methods , Radioimmunoassay/statistics & numerical data
14.
J Infect Dis ; 171(3): 632-8, 1995 Mar.
Article in English | MEDLINE | ID: mdl-7876610

ABSTRACT

The safety and immunogenicity of a group A plus group C meningococcal polysaccharide-CRM197 conjugate vaccine was evaluated in 304 8- to 10-week-old Gambian infants. Infants were immunized with one, two, or three doses of conjugate vaccine or with two doses of a meningococcal A plus C polysaccharide vaccine. The conjugate vaccine produced few systemic side effects, and local reactions were similar to those produced by the polysaccharide vaccine. Postvaccination group A meningococcal polysaccharide antibody levels, measured by ELISA, increased progressively after one, two, or three doses of conjugate vaccine. However, one dose of conjugate vaccine given at the age of 6 months induced a higher group C meningococcal antibody response than did two doses of conjugate vaccine given at 2 and 6 months. Two doses of conjugate vaccine induced higher levels of antibody than did two doses of polysaccharide vaccine. Thus, this new meningococcal conjugate vaccine proved to be safe and immunogenic.


Subject(s)
Bacterial Proteins/immunology , Bacterial Vaccines/immunology , Neisseria meningitidis/immunology , Polysaccharides, Bacterial/immunology , Antibodies, Bacterial/blood , Bacterial Vaccines/adverse effects , Female , Humans , Infant , Male , Meningococcal Vaccines , Vaccination , Vaccines, Conjugate/immunology
15.
Clin Diagn Lab Immunol ; 2(2): 132-7, 1995 Mar.
Article in English | MEDLINE | ID: mdl-7697519

ABSTRACT

A new standard meningococcal reference serum designated CDC1992 was prepared to replace meningococcal reference sera ECG and PB-2, which are not available in sufficient quantities for continued use as primary reference sera. CDC1992 was prepared from 14 healthy adult volunteers who underwent plasmapheresis 4 to 12 weeks postvaccination with a single dose of a Neisseria meningitidis quadrivalent polysaccharide vaccine. Total and/or class-specific meningococcal serogroup A and C anticapsular antibody concentrations (in micrograms per milliliter) were assigned to CDC1992 by using homologous and heterologous enzyme-linked immunosorbent assay (ELISA) formats. The reference serum ECG was used as a reference standard to assign total anticapsular antibody concentrations to CDC1992 by a homologous ELISA format. A heterologous ELISA format, with the Haemophilus influenzae type b standard reference serum FDA 1983, was used to assign total and class-specific antibody concentrations to CDC1992. Alkaline phosphatase-labeled mouse anti-human monoclonal antibody conjugates were used as secondary antibodies in both ELISA formats. The total, immunoglobulin G (IgG), IgA, and IgM antibody concentrations, assigned to CDC1992 for serogroup A were 135.8, 91.8, 20.1, and 23.9 micrograms/ml, respectively, and those for serogroup C were 32.0, 24.1, 5.9, and 2.0 micrograms/ml, respectively. Meningococcal serogroup A and C antibody concentrations were in good agreement when homologous and heterologous ELISA format results were compared. Total and class-specific serogroup A and C antibody concentrations were determined in six adult quality control serum samples from the Centers for Disease Control and Prevention by using the homologous ELISA and our assigned antibody concentrations for CDC1992. Antibody concentrations in reference sera ECG and PB-2 were measured in order to provide a historical link to previous studies. The general acceptance of CDC1992 as the standard reference serum and the assigned antibody concentrations will allow investigators to compare antibody levels in serum to those in a single reference preparation.


Subject(s)
Antibodies, Bacterial/analysis , Bacterial Capsules/immunology , Neisseria meningitidis/classification , Neisseria meningitidis/immunology , Serology/standards , Adult , Antibody Specificity , Enzyme-Linked Immunosorbent Assay , Humans , Reference Standards , Vaccination
16.
J Clin Microbiol ; 32(10): 2441-7, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7814480

ABSTRACT

There is a lack of consensus among investigators who use a variety of immunoassay techniques (e.g., enzyme-linked immunosorbent assay [ELISA] and radioimmunoassay) regarding the protocols for describing and forming standard reference or calibration curves and interpolating serum antibody concentrations. This confounds the issue of detecting the presence or absence of parallelism between standard reference serum and serially diluted serum sample curves. These curves must be parallel to support the assumption that the antibody-binding characteristics are similar enough to allow the determination of antibody levels in the diluted serum sample. There is no universal and widely adopted strategy for assessing parallelism in bioassays, and without an assurance of parallelism, investigators are not able to calculate reliable estimates for antibody concentrations in serum samples. Furthermore, single-point (dilution) serum assays do not provide information related to parallelism and nonparallelism, and this, too, may lead to considerable error when calculating antibody concentrations. When assay methodology, technique, and precision improve to the extent that standard reference serum and serially diluted serum sample curves are fit with little error, standard analysis of variance techniques are overly sensitive to negligible departures from parallelism. We present a series of guidelines that compose a protocol for assessing parallelism between bioassay dilution curves that are applicable to data derived from ELISAs. These criteria should be applicable, with minor modifications, to most immunoassay experimental situations and, most importantly, are not dependent on the mathematical model used to form the standard reference curve. These guidelines have evolved in our laboratories over the past 4 years during the performance of thousands of ELISAs for antibodies to the capsular polysaccharides of Neisseria meningitidis groups A and C and Haemophilus influenzae type b.


Subject(s)
Enzyme-Linked Immunosorbent Assay/standards , Analysis of Variance , Antibodies, Bacterial/blood , Haemophilus influenzae/immunology , Humans , Neisseria meningitidis/immunology
17.
Infect Immun ; 62(8): 3391-5, 1994 Aug.
Article in English | MEDLINE | ID: mdl-8039909

ABSTRACT

A meningococcal vaccine containing group A and C polysaccharides conjugated to CRM197 was evaluated in 50 adults. Vaccinees were entered into one of five groups: 30 adults received a single dose of either 22, 11, or 5.5 micrograms of the conjugated A-C vaccine; 10 received an approved meningococcal vaccine; and 10 received saline injections. Local and systemic reactions to vaccines were recorded, and immune responses were determined. The experimental meningococcal vaccine was well tolerated, with the most frequent reaction being pain at the injection site. Both A and C polysaccharide components of the experimental vaccine were highly immunogenic, and total antibody concentrations 1 month postvaccination were not significantly different from the mean antibody concentrations among adults given the approved meningococcal vaccine. In addition, significant rises in immunoglobulin G, A, and M antibodies to both A and C polysaccharides occurred. Antibody concentrations measured at 6 and 12 months postvaccination had declined but remained significantly higher than prevaccination concentrations. Postvaccination meningococcal group C functional antibody activity increased more than 600-fold for both the polysaccharide and the conjugate vaccines. Further studies of this conjugated meningococcal vaccine are indicated for young children and infants.


Subject(s)
Antibodies, Bacterial/blood , Bacterial Vaccines/immunology , Neisseria meningitidis/immunology , Polysaccharides, Bacterial/immunology , Adolescent , Adult , Bacterial Vaccines/adverse effects , Blood Bactericidal Activity , Humans , Meningococcal Vaccines , Middle Aged , Vaccines, Conjugate/immunology
18.
J Clin Microbiol ; 32(6): 1475-82, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8077392

ABSTRACT

A standardized enzyme-linked immunosorbent assay (ELISA) was used by 11 laboratories to measure levels of total serum antibody to Neisseria meningitidis serogroup C capsular polysaccharide in 16 unpaired pre- and postvaccination serum samples. Twelve serum samples were from adults, and four were from children aged 2, 3, 5, and 9. The between-laboratory coefficient of variation for pre- and postvaccination sera ranged from 16 to 59% and 11 to 21%, respectively. The average percent difference (absolute value) from the between-laboratory means for all prevaccination sera measured by each laboratory was 24%, whereas the average percent difference was 13% for all postvaccination sera. A postvaccination quality control serum was diluted three times to give optical densities on the high, middle, and low portions of the standard reference curve. The three dilutions were assayed by the 11 laboratories a total of 241 times and yielded an overall coefficient of variation of 20%. Antibody-binding inhibition curves showed that the standardized ELISA was specific for N. meningitidis serogroup C capsular polysaccharide antibody. Fifty percent inhibition of seven serum samples was obtained after reaction with an average concentration of 0.9 micrograms of meningococcal serogroup C polysaccharide per ml; an average of 93% inhibition was obtained with 50 micrograms of polysaccharide per ml. The acceptance and use of this standardized ELISA will reduce between-laboratory assay variability and ensure a more accurate and reproducible assessment of immunogenicity for vaccines under development.


Subject(s)
Antibodies, Bacterial/blood , Enzyme-Linked Immunosorbent Assay , Neisseria meningitidis/immunology , Polysaccharides, Bacterial/immunology , Adult , Antibodies, Bacterial/immunology , Child , Child, Preschool , Humans , Neisseria meningitidis/classification , Reference Standards , Reproducibility of Results , Vaccination
19.
J Am Mosq Control Assoc ; 10(1): 14-23, 1994 Mar.
Article in English | MEDLINE | ID: mdl-8014623

ABSTRACT

Since a 1988-89 survey of northern New Zealand revealed no additions to the known mosquito fauna, this country's used tire importations have much increased. Relevant entomological quarantine was thus monitored in a November 1992-January 1993 Auckland project, during which almost 1/3 of 8,549 casings from Japan proved wet on inspection. In this study and at 2 South Island ports afterwards, 5 vessels from Japan and one from Australia were found to have brought in mosquito-infested used tires. Live Aedes albopictus (all larval instars, pupae, and adults) and Aedes japonicus, and dead Tripteroides bambusa were discovered in shipments from Japan (3 interceptions each in the first 2 cases, and one in the 3rd). Live Tripteroides tasmaniensis were recorded from the Australian cargo. One of the Ae. albopictus arrivals was followed by an apprehended introduction at an Auckland importer's premises.


Subject(s)
Aedes , Commerce , Mosquito Control/methods , Animals , Automobiles , Ecology , New Zealand , Population Control , Population Surveillance , Species Specificity , Water
20.
Diagn Cytopathol ; 9(6): 691-7, 1993 Dec.
Article in English | MEDLINE | ID: mdl-8143548

ABSTRACT

All fine-needle aspirates (FNA) performed on the male breast at The University of Texas M. D. Anderson Cancer Center from 1985 to 1992 were reviewed, totaling 64. The patients' ages ranged from 19 to 86 years, with a mean of 56 years. Thirty-three patients had a history of an extramammary malignancy. The diagnoses established by FNA were gynecomastia (45), mammary carcinomas (6), neoplasms metastatic to the breast (5), suspicious for carcinoma (1), intra-mammary lymph node (1), and lipoma (1). In five cases the aspirates were nondiagnostic. Two of these proved to be gynecomastia on subsequent histologic examination. Of the six FNA cases initially thought to represent primary breast carcinomas, two were found to be secondary because of involvement of the underlying chest wall by mesothelioma (1), and mucinous adenocarcinoma, unknown primary (1). No false-positive diagnosis was rendered. We conclude that fine-needle aspiration of the male breast is a reliable means of assessment; however, unique problems may be encountered compared with aspiration of the female breast. These include the epithelial hyperplasia frequently associated with gynecomastia, the relatively equal frequency of primary and metastatic breast lesions when a malignant process is discovered, and chest wall lesions masquerading as breast lesions.


Subject(s)
Breast Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Biopsy, Needle , Breast Neoplasms/secondary , Cancer Care Facilities , Gynecomastia/pathology , Humans , Male , Middle Aged , Neoplasm Metastasis , Retrospective Studies
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