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Results for “Bioactivation”

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At least 289 records · Page 16Linked to original sources

PubMed1997

Leucocyte-derived bioactive substances in fresh frozen plasma.

Bioactive substances in fresh frozen plasma (FFP) are considered to be related to adverse reactions after transfusion, particularly in septic or traumatized patients. Therefore, we analysed the concentration of various bioactive substances (histamine, eosinophil cationic protein, eosinophil protein X, myeloperoxidase and interleukin-6) in 25 u. of thawed FFP from healthy donors. These were compared with donor plasma concentrations of 24 healthy controls. In addition, we analysed the concentration of the bioactive substances, except interleukin-6, in 25 u. of thawed FFP, which were subjected to leucocyte filtration before freezing and storage. Finally, we analysed the substances in 10 leucocyte non-filtered plasma units before freezing and storage, and after thawing, respectively. Before analyses, which were performed by ELISA and RIA methods, these latter samples were sterile filtered through a 0.20-micron filter. Histamine, eosinophil cationic protein, eosinophil protein X and myeloperoxidase concentrations were significantly greater (P < 0.05) in the 25 u. of FFP compared with normal donor plasma. Pre-storage leucocyte filtration reduced concentrations of the bioactive substances in FFP to concentrations comparable with normal donor plasma concentrations. Interleukin-6 was undetectable in all FFP units and in 21 of the 24 control donors. Histamine, eosinophil cationic protein and myeloperoxidase concentrations were significantly higher (P < 0.05) in samples collected from the 10 u. of FFP after freezing and thawing compared with samples collected before freezing. We conclude that fresh frozen plasma prepared by a conventional separation method contains various leucocyte-derived bioactive substances, which may be reduced by pre-storage leucocyte filtration.

Blood Preservation↗

PubMed1991

Roles of different cytochrome P450 enzymes in bioactivation of the potent hepatocarcinogen 3-methoxy-4-aminoazobenzene by rat and human liver microsomes.

The potent hepatocarcinogen 3-methoxy-4-aminoazobenzene (3-MeO-AAB) has been reported to be bioactivated to mutagenic intermediates by rat liver microsomal cytochrome P450 (P450) and to be a selective inducer of rat P450IA2. In this study we have further investigated the roles of individual rat and human P450 enzymes in the bioactivation of this hepatocarcinogen in a Salmonella typhimurium TA1535/pSK1002 system where umu response is indicative of DNA damage. 3-MeO-AAB was found to be bioactivated by liver microsomal enzymes from rats and humans in this assay system. The liver microsomal activities are increased by pretreatment of rats with various P450 inducers such as phenobarbital (PB), beta-naphthoflavone (BNF), dexamethasone (DEX), acetone, ethanol, isoniazid (INH), diphenylhydantoin and valproic acid, and can be inhibited considerably by SKF-525A and metyrapone. alpha-Naphthoflavone (ANF) is also an inhibitor for the reaction catalyzed in BNF-treated rats, but stimulated the microsomal activity in DEX-treated rats. Evidence has also been obtained that specific antibodies raised against P450IIB1, P450IA1 or IA2, P450IIE1, and P450IIIA2 inhibited the activation in liver microsomes from rats pretreated with PB, BNF, INH and DEX respectively, suggesting the possible roles of several P450 enzymes in the bioactivation of 3-MeO-AAB. The results obtained with reconstituted monooxygenase systems containing various rat P450 enzymes are highly supportive of this conclusion. Human liver microsomal activation of 3-MeO-AAB was also inhibited to various extents by antibodies raised against P450IA2, P450MP, P450IIE1 and P450IIIA4. In a reconstituted system containing purified forms of human P450, P450IA2 was the most active in catalyzing 3-MeO-AAB, followed by P450IIIA4 and P450MP. ANF, a known activator of P450IIIA-catalyzed reactions, caused an increase in activation of 3-MeO-AAB in human liver microsomal and P450IIIA4- and P450MP-containing reconstituted systems. From these results it is concluded that multiple P450 enzymes in rat and human liver microsomes are involved in the bioactivation of 3-MeO-AAB, regardless of its selective induction of the rat P450IA2 gene.

Animals↗

PubMed1995

Gonadotrophin surge-attenuating factor bioactivity is present in follicular fluid from naturally cycling women.

Rat pituitary monolayer bioassays were used to compare gonadotrophin surge-attenuating factor (GnSAF) bioactivity in follicular fluid from 12 follicles in 10 spontaneously cycling women with that in pooled follicular fluid from women undergoing ovulation induction. Expressed as ED50S (microliter follicular fluid/well producing 50% of maximal effect), GnSAF bioactivity was detectable in all spontaneous follicular fluid samples (1.4-33.3 microliters/well) and in follicular fluid from women undergoing ovulation induction (6.8 microliters/well). This GnSAF bioactivity was unaffected by pre-incubation with an inhibin antibody. When the data were grouped according to whether the recovered oocytes fertilized in vitro or not, the fertilized group contained significantly greater GnSAF bioactivity than the unfertilized group (5.3 +/- 1.1 and 14.1 +/- 2.6 microliters/well respectively, P < 0.05). While both inhibin bioactivity (9.7 +/- 1.4 and 28.9 +/- 12.1 microliters/well) and immunoreactivity (36.8 +/- 2.2 and 21.0 +/- 3.0 and ng/ml) were also greater (P < 0.01) in the fertilized compared with the unfertilized groups respectively, there were no other significant differences between the two groups. We conclude that GnSAF is found in follicular fluid from spontaneously cycling women, supporting in-vivo evidence for the involvement of GnSAF in feedback control of the ovary-pituitary axis.

Adult↗

PubMed1998

Immunoneutralization of follicle stimulating hormone does not affect gonadotrophin surge-inhibiting factor/attenuating factor bioactivity during the rat ovarian cycle.

The physiological role of follicle stimulating hormone (FSH) in the regulation of the release of the putative ovarian factor gonadotrophin surge-inhibiting or -attenuating factor (GnSIF/AF) was investigated. Blood FSH concentrations were immunoneutralized in female rats during different days of the ovarian cycle. FSH antiserum was injected in different protocols to neutralize the FSH surge(s) and/or basal FSH concentrations. All animals were killed on the morning of the day of pro-oestrus of the control rats. Hemi-pituitaries were incubated and gonadotrophin-releasing hormone (GnRH) stimulated and basal luteinizing hormone (LH) and FSH releases and pituitary contents were measured. The biological effectiveness of the FSH antiserum was established by monitoring the histology of vaginal smears and inhibin concentrations in trunk-blood. The predominance of small leukocytes in the smears (low oestradiol bioactivity) and the decreased (bioactive) inhibin secretion imply that anti-FSH neutralizes FSH bioactivity. Simultaneously, the results showed that all anti-FSH injections did not affect the biphasic LH response to GnRH in vitro, which shows an unaltered presence of GnSIF/AF bioactivity in the blood circulation. The results suggest that under physiological conditions GnSIF/AF bioactivity, which keeps the LH responsiveness of the pituitary gland suppressed to the action of GnRH, is not supported by inhibin and not controlled by FSH.

Animals↗

PubMed2000

Choriocarcinoma-like human chorionic gonadotrophin (HCG) and HCG bioactivity during the first trimester of pregnancy.

The objective of this study was to evaluate the distribution of choriocarcinoma-like human chorionic gonadotrophin (HCG) isoforms during first trimester pregnancy and their relationship with in-vitro HCG bioactivity. This was done by means of a retrospective analysis of patients' sera with first trimester normal intrauterine and abnormal (ectopic) pregnancies. Serum samples were obtained from 38 women with an amenorrhoea of <10 weeks. From these, 19 had a normal intrauterine pregnancy (IUP) and 19 an ectopic pregnancy (EP). Total immunoreactive HCG (HCGi), free beta-HCGi and oestradiol were measured by enzyme immunoassays and bioactive HCG by the mouse Leydig cell bioassay. The alterations in HCG isoform content were measured by the combination of two immunometric assays, B152 for choriocarcinoma-like HCG and B109 for intact HCG detection and expressed as the B152/B109 ratio. Choriocarcinoma-like HCG isoforms ratio measured by B152 and B109 assays was significantly higher in the low subgroups of free beta-HCGi and gestational age (P = 0.0111 and 0.0036 respectively). Whereas bioactive to immunoreactive HCG ratios (b/i ratio) were significantly higher when free beta-HCGi concentrations were low (P = 0.0010), no correlation was found between the variation of bioactivity (b/i ratio) and the proportion of choriocarcinoma-like HCG isoforms (B159/B108). It is concluded that in first trimester pregnancies (i) the modulation of HCG in-vitro bioactivity is not related to the variation of choriocarcinoma-like HCG isoforms secretion and (ii) the amount of choriocarcinoma-like HCG isoforms secreted by the early trophoblast is predominant and may be the result of an early developmental regulation of glycosylation enzyme.

Amenorrhea↗

PubMed1996

Soluble tumor necrosis factor (TNF) receptors conserve TNF bioactivity in meningitis patient spinal fluid.

The content of tumor necrosis factor (TNF)-alpha antigen and the bioactivity of soluble TNF receptor type II (sTNF-RII) in cerebrospinal fluid (CSF) from 29 patients with meningeal symptoms and fever were examined. Immunoreactive TNF was demonstrated in CSF from 4 of 7 patients with bacterial meningitis. In 3 of 8 patients with aseptic meningitis, CSF also contained TNF, but TNF bioactivity was confined to samples from patients with bacterial meningitis. Bioactive TNF was exclusively in high-performance liquid chromatography fractions containing 30- to 60-kDa proteins. Lipopolysaccharide induced down-regulation, possibly after shedding of granulocyte surface membrane TNF-RII. Consistently, there was a statistically significant correlation between sTNF-RII and CSF leukocyte counts. Bioactive TNF was found only in CSF containing >1 ng of sTNF-RII/mL; samples without TNF bioactivity contained less sTNF-RII. Thus, a stabilizing effect of sTNF-RII on the oligomeric cytokine in vivo is plausible.

Cells, Cultured↗

PubMed2002

A 60-66 kDa protein with gonadotrophin surge attenuating factor bioactivity is produced by human ovarian granulosa cells.

We aimed to confirm the ovarian site of gonadotrophin surge-attenuating factor (GnSAF) production and produce granulosa/luteal cell-conditioned medium (G/LCM) containing GnSAF for purification studies. Blue dye affinity chromatography followed by pseudochromatofocusing of G/LCM yielded bioactive fractions at pH 5.74 and 5.77. The former had a major 60-66 kDa band with an internal amino acid sequence of EPQVYVHAP following tryptic digestion. A rat polyclonal antiserum (rPAb) raised against this band completely blocked in-vitro GnSAF bioactivity in human follicular fluid, serum and G/LCM. GnSAF bioactivity was localized to a 64 kDa band of serum-free G/LCM and following 2D gel electrophoresis, one of the spots recognized by Western blotting with the GnSAF rPAb had an N-terminal amino acid sequence of NH-XVPQGNAXXN. Neither amino acid sequence had significant homology with proteins in the human genome database. When ovarian tissues from spontaneously cycling women were cultured under serum-free conditions, neither theca- nor stroma-conditioned media contained GnSAF bioactivity. However, granulosa cell-conditioned medium significantly reduced GnRH-induced LH secretion, an effect that was reversed by incubation with the GnSAF rPAb. In conclusion, we have confirmed that human granulosa cells produce GnSAF within the ovary and have two candidate amino acid sequences for GnSAF. We have also demonstrated that serum-free granulosa cell culture constitutes the method of choice for the characterization of GnSAF since recovery of bioactivity is superior in the presence of fewer serum proteins.

Biological Factors↗

PubMed1992

Changes in peripheral levels of bioactive and immunoreactive inhibin, estradiol-17 beta, progesterone, luteinizing hormone, and follicle-stimulating hormone associated with follicular development in cows induced to superovulate with equine chorionic gonadotropin.

Changes in concentrations of bioactive and immunoreactive (ir-) inhibin, estradiol-17 beta, progesterone, LH, and FSH in peripheral blood were determined in cows induced to superovulate with eCG. The pattern of follicular growth was also characterized by daily ultrasonographic examination. Hormonal profiles and follicular development during the intact estrous cycle of the same animals before eCG treatment served as controls. Equine CG increased the number of follicles of various sizes (small, greater than or equal to 4 less than 7, medium, greater than or equal to 7 less than 10; large, greater than or equal to 10 mm in diameter) by 4 days after administration. The second growth of large follicles occurred within 1 day after superovulation. Inhibin bioactivity in jugular vein blood was detectable 48 h after eCG injection (44 h before LH peak), whereas it was not detected before administration of eCG or during control cycles. Circulating levels of bioactive inhibin further increased during the two waves of growth of large follicles. The highest activity of inhibin was noted at the time of the preovulatory LH peak (0 h). Thereafter, bioactivity of inhibin in peripheral plasma dropped from 0 to 24 h after the LH peak, and the activity increased again at 72 h compared to the value at -44 h. Plasma levels of ir-inhibin showed a pattern similar to changes in bioactive inhibin in the eCG-treated cows. Plasma concentrations of estradiol-17 beta also increased concomitantly with two waves of growth of large follicles. There was no correlation between plasma levels of progesterone and inhibin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Luminescence luteinizing hormone/choriogonadotropin (LH/CG) bioassay: measurement of serum bioactive LH/CG during early pregnancy in human and macaque.

Because of the microheterogeneities of gonadotropins, measurement of immunoreactivity of these glycoproteins does not necessarily reflect changes in their bioactivity. In addition, LH bioactivities in human samples analyzed by a rodent LH bioassay have been discordant with findings based on human granulosa-luteal cells. We have isolated a human LH/choriogonadotropin (CG) receptor cDNA and expressed the recombinant protein. Using 293 cells permanently transfected with the human LH receptor cDNA and a luciferase reporter gene driven by a cAMP-dependent promoter, we have developed a luminescence LH/CG bioassay. After cells were treated with human LH or CG for 20 h, luciferase activity was measured through use of a luminometer. Luciferase activity in the cells was increased in a dose-dependent manner. In contrast, treatment with FSH, thyroid-stimulating hormone, prolactin, growth hormone, adrenocorticotropin, insulin, prostaglandins, and several neurotransmitters had no effect. Because treatment with basic fibroblast growth factor (bFGF) caused significant increases in basal luciferase activity, a fixed amount of bFGF was included in all reactions. Incubation with 0.1 to 30 microliters serum from women during different physiological states stimulated the luciferase activity in parallel with the hCG standard curve. In 4 conception cycles, bioactive LH/hCG levels began to increase 2 wk after the midcycle LH surge, followed by a logarithmic increase from 22 days on. Due to the lack of a homologous RIA for measuring CG levels in monkeys, we analyzed serum bioactive monkey CG (mCG) in macaque during early pregnancy. Bioactive mCG was detected about 12 days after the midcycle LH surge and fertile mating and persisted until Days 21-23, followed by a decline.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1995

Gonadotropin surge-attenuating factor bioactivity in serum from superovulated women is not blocked by inhibin antibody.

Primary pituitary cultures from adult female rats were used to investigate gonadotropin surge attenuating factor (GnSAF) bioactivity. Serum from superovulated women was charcoal-treated and incubated with either an inhibin antibody (MC4), normal sheep serum (NSS), or serum-free defined medium (SFDM) before addition to cell culture. The reduction in GnRH-induced LH secretion (GnSAF bioactivity) produced by the serum (30.1 +/- 6.5%, p < 0.001, of control) was not altered by prior incubation with either MC4 or NSS (24.9 +/- 3.6 and 23.2 +/- 2.7%, p < 0.001, of control, respectively). This indicates that GnSAF bioactivity present in serum from superovulated women is not entirely attributable to inhibin. Recombinant human inhibin reduced basal FSH secretion to 24.6 +/- 4.7% (p < 0.001) of the control value. However, this was totally abolished by prior incubation with MC4, but not NSS. We have also shown that ovarian steroids are not responsible for GnSAF bioactivity in vitro. In conclusion, in contrast to findings in superovulated rats, inhibin antibody did not block GnSAF bioactivity in serum from superovulated women.

Animals↗

PubMed2002

Holding power of bioabsorbable ciprofloxacin-containing self-reinforced poly-L/DL-lactide 70/30 bioactive glass 13 miniscrews in human cadaver bone.

Antibiotics-plus bioactive glass-containing bioabsorbable self-reinforced (SR) polylactide screws have been developed for antibacterial osteoconductive bone fixation. The aim of the present study was to test the pullout properties of these recently developed miniscrews. Ciprofloxacin-plus bioactive glass-containing SR-polylactide miniscrews (BC) were compared with miniscrews made of neat SR-polylactide (A), SR-polylactide with bioactive glass (B), and ciprofloxacin-containing SR-polylactide (C). BC miniscrews and their controls (A, B, C) (all of length 6.0 mm, core diameter 1.45 mm, thread diameter 2.0 mm) were applied to one pair of cadaveric fibulae. Pullout force was measured using a materials testing machine. We carried out 49-50 pullout tests for each implant type. The Mann-Whitney test and Student's t-test were used for statistical evaluation. The pullout force for BC miniscrews was 114.9 +/- 34.0 (SD) N. Pullout forces for control miniscrews were 162.7 +/- 37.8 N (A), 99.1 +/- 16.2 N (B), and 142.9 +/- 26.9 N (C). Differences between the four groups were statistically significant (p < 0.001). Ciprofloxacin-plus bioactive glass-containing polylactide miniscrews have good holding power to human cadaver fibulae. However, adding bioactive glass and ciprofloxacin components to neat SR-polylactide results in lower pullout values.

Absorbable Implants↗

PubMed1997

Effect of heating on extracellular bioactive substances in stored human blood: in vitro study.

BACKGROUND: We have previously shown extracellular accumulation of various leukocyte and platelet-derived bioactive substances in human blood during storage. Release of bioactive substances may be temperature-dependent, and we studied the effect of heating during in vitro transfusion on bioactive substance accumulation in stored human blood. METHODS: Eight units of whole blood and eight units of prestorage leukofiltered whole blood were stored at 4 degrees C for 7 days. Subsequently, the blood from all 16 units was transfused via a blood-heating device, which increased the blood temperature to 37 degrees C at outlet. Samples for enzyme-linked immunosorbent assay or radioimmunoassay analyses of histamine, myeloperoxidase (MPO), eosinophil cationic protein (ECP), and plasminogen activator inhibitor-1 (PAI-1) were drawn from the units at donation, after 7 days of storage just before transfusion, and during the in vitro transfusion. RESULTS: Extracellular concentrations of histamine, MPO, ECP, and PAI-1 were significantly (p < 0.05) increased in nonfiltered whole blood stored for 7 days compared with concentrations in fresh donated blood and in prestorage leukofiltered whole blood stored for 7 days. Heating reduced histamine, MPO, and ECP concentrations significantly (p < 0.05) in nonfiltered whole blood, whereas PAI-1 concentrations increased significantly (p < 0.05). Finally, there was no difference in concentrations of histamine, MPO, ECP, and PAI-1 in samples collected before and after heating of leukofiltered whole blood. CONCLUSIONS: Heating reduces accumulation of extracellular leukocyte-derived bioactive substances in whole blood, whereas it increases platelet-derived substances. Prestorage leukofiltration, however, reduces the obligatory extracellular accumulation of leukocyte and platelet-derived bioactive substances, which in addition is unchanged by heating.

Blood Preservation↗

PubMed2001

Bioactive bone cement as a principal fixture for spinal burst fracture: an in vitro biomechanical and morphologic study.

STUDY DESIGN: An in vitro biomechanical and radiographic study to evaluate the properties of a newly developed bioactive bone cement for stabilization of the fractured spine, suitable for minimally invasive application. OBJECTIVES: To determine the mechanical stability of the fractured spine after injection of the newly developed bioactive bone cement under quasi-static and cyclic loading regimens. SUMMARY OF BACKGROUND DATA: Bone cement injection has been reported as a potentially useful, minimally invasive technique for treating vertebral body fracture or stabilizing osteoporosis. However, potential problems associated with the use of polymethylmethacrylate (PMMA) have prompted the search for alternative solutions. The use of bioactive bone cement as a potential replacement for PMMA has been reported. METHODS: Biomechanical and radiographic analyses were used to test the mechanical stability of the fractured spine. The cement used was formed from hydroxyapatite powder containing strontium and bisphenol A diglycidylether dimethacrylate (D-GMA) resin. Twenty-six fresh porcine spine specimens (T10-L1) were divided into three groups: pilot, intact, and cemented. Spinal stiffness and failure strength were recorded in the intact group with the specimens flexed at 10 degrees. Uniform injuries were created in all specimens of the cemented group, and compressive loading was applied with 10 degrees of flexion until a fracture occurred. The bone cement was injected into the fractured spine, and stiffness was evaluated after 1 hour. Failure strength was also recorded after 3000 and 20,000 fatigue load cycles. Morphology of the specimens was observed and evaluated. RESULTS: Results from a cell biocompatibility test indicated that the new bioactive bone cement was favorable for cell growth. Spinal stiffness significantly decreased after fracture (47.5% of intact condition). Instant stiffness of the spine recovered to 107.8% of the intact condition after bone cement injection. After 3000 and 20,000 cycles of fatigue loading, stiffness of the cemented spine was found to be 93.5% and 94.4% of intact stiffness, respectively (P < 0.05). Average failure strength of the spine was 5056 N (after 3000 cycles) and 5301 N (after 20,000 cycles) after bone cement injection and fatigue loading. Radiographs and cross-sectional observations indicated a good cement-bone bonding and fracture fill. CONCLUSIONS: A new bioactive bone cement without cytotoxic effect has been developed. Results show that minimally invasive techniques to apply this cement to porcine spines results in augmentation of mild burst fractures such that the original stiffness and strength of the vertebra are recovered. This new cement therefore shows potential as an augmentation to traditional instrumentation in the surgical management of vertebral fractures. The potential for further clinical applications is currently under investigation.

Animals↗

PubMed1997

Bone remodeling with resorbable bioactive glass and hydroxyapatite.

Particulate resorbable bioactive glass and resorbable hydroxyapatite did not inhibit the growth of periodontal ligament cells and osteoblasts in cultures. Both cell types promoted the activation of alkaline phosphatase around the surface of bioactive glass particles. When bioactive glass and hydroxyapatite were cultured with peripheral blood lymphocytes/monocytes, the number of nonadherent and adherent cells was the same as observed with a control. Resorbable bioactive glass activated the cells necessary for bone formation better than hydroxyapatite and suppressed the formation of osteoclasts for bone resorption. Resorbable bioactive glass and resorbable hydroxyapatite were found to be harmless to periodontium cells and lymphocytes/monocytes.

Absorption↗

PubMed2006

Bioactivity of postshock mesenteric lymph depends on the depth and duration of hemorrhagic shock.

Mesenteric hypoperfusion due to circulatory shock is a key event in the pathogenesis of subsequent distant organ injury. Postshock mesenteric lymph (PSML) has been shown to contain proinflammatory mediators elaborated from the ischemic gut. We hypothesize that the relative bioactivity of PSML depends on the depth and duration of circulatory shock. To first determine the timing of PSML bioactivity, we subjected rats to hemorrhagic shock (30 mm Hg x 45 min) and then resuscitation with 50 vol% of shed blood and normal saline (4x shed blood) over 2 h. Mesenteric lymph was collected hourly up to 6 h after shock. Superoxide release was measured from human neutrophils (polymorphonuclear neutrophils [PMNs]) incubated with lymph fractions collected from each of the hourly time points. Rats were then subjected to four different shock variations: (1) 30 mm Hg x 45 min, (2) 30 mm Hg x 15 min, (3) 45 mm Hg x 45 min, and (4) 45 mm Hg x 15 min, and were resuscitated. PSML flow depends on depth of shock, but not duration of shock or resuscitation volume. Maximal PSML bioactivity, as measured by PMN priming for the respiratory burst, occurred during the third postshock hour, which correlated with peak lymph flow rate. PSML bioactivity was greatest with 30 mm Hg x 45 min, followed by 30 mm Hg x 15 min, 45 mm Hg x 45 min, and 45 mm Hg x 15 min. Hemorrhagic shock provokes the release of bioactive agents in PSML that is dependent on both depth and duration of shock.

Animals↗

PubMed1994

Serum lactogens possessed normal bioactivity in patients with lactation insufficiency.

OBJECTIVE: Insufficient breast milk is the most common reason for premature termination of breast-feeding. The causes of lactation insufficiency are usually multifactorial; in a small percentage of cases it is due to primary lactation failure of unknown origin. The aim of this study was to investigate whether lactation insufficiency of unknown origin could be caused by serum lactogens that had reduced biological activity. DESIGN: Women with lactation insufficiency of unknown origin and normal lactating controls were subjected to a standardized breast-feeding test for assessment of milk production. Thirty minutes later, serum samples were obtained for determination of total lactogen bioactivity, using an in-vitro bioassay, and levels of prolactin (PRL) and growth hormone (GH) using radioimmunoassay (RIA). PATIENTS: Twelve lactating mothers with a clinical diagnosis of lactation insufficiency of unknown origin were compared with 12 matched mothers with normal lactation. MEASUREMENTS: The Nb2 lymphoma cell bioassay was used to measure total lactogen bioactivity in sera. Conventional RIA kits were used to estimate serum PRL and GH concentrations. RESULTS: Mean milk yield on standardized test feed was 21.6 ml for patients and 146.5 ml for controls. In both patient and control groups the total serum lactogen bioactivity ranged from about 150 to 5000 mIU/l, while the serum RIA (PRL+GH) levels ranged from about 350 to over 7000 mIU/l. There was no evidence of lactogens with reduced bioactivity in the patients' sera. CONCLUSION: Lactation insufficiency in the women studied cannot be explained by serum lactogens that possess unusually low bioactivity.

Biological Assay↗

PubMed1995

Age related changes in the pituitary-testicular axis in normal men; lower serum testosterone results from decreased bioactive LH drive.

OBJECTIVE: The mechanism underlying the slight hypoandrogenism associated with normal ageing remains unclear. We have therefore examined age related changes in the activity of the pituitary-testicular axis in healthy normal males. DESIGN: Random blood samples were obtained from 219 normal men whose ages ranged from 20 to 79 years. At the time of the study, none of the men had received treatment or had ever had any endocrine dysfunction diagnosed. MEASUREMENTS: Luteinizing hormone was measured in the subjects' plasma using a commercially available immunoradiometric assay (IRMA, Serono Maiaclone) and a fully validated in-vitro bioassay. Testosterone and FSH were measured using standard radioimmunoassays (RIA) whilst sex hormones binding globulin was assayed by an IRMA. RESULTS: Levels of total testosterone (total-T) and bioactive LH fell with age (r = -0.231 and -0.189 respectively) by 5.9 nmol/l and 2.3 IU/l respectively between grouped patients aged 20-39 years (Group A) and 60-79 years (Group C). In contrast, immunoreactive LH remained unchanged. The LH B:I ratio also fell with age (P < 0.0001) being 5.0 +/- 0.3 (group A) and 3.3 +/- 0.2 (group C), representing a fall of 33%. Since immunoreactivity remained constant, this fall primarily represented the decline in LH bioactivity. Bioactive, but not immunoreactive LH correlated to total-T (P = 0.009, n = 209) and the total-T:LH ratios fell by over 30% between groups A and C using the IRMA, but remained unchanged by bioassay. CONCLUSIONS: There is an underlying decline in both total-testosterone and free-testosterone index, and bioactive LH levels with advancing age, suggestive of a hypothalamo-pituitary defect which leads to lower bioactive LH levels which in turn are responsible for the diminished gonadal steroidogenesis. Elucidation of the mechanism underlying this slight decline in hypothalamopituitary testicular activity is complicated by differences between the data obtained by immunoassay or bioassay.

Adult↗

PubMed1990

Measurement of tissue cholecystokinin (CCK) concentrations by bioassay and specific radioimmunoassay: characterization of the bioactivity of CCK-58 before and after tryptic cleavage.

Cholecystokinin (CCK) was measured in an extract of porcine brain by a bioassay system based on the enzyme release from isolated pancreatic acini and a specific radioimmunoassay. The tissue extract represented a crude peptide preparation of porcine brain that contained several molecular forms of CCK, most of them of high molecular weight including CCK-58. The different molecular forms of CCK in the porcine brain extract were isolated by high performance liquid chromatography (HPLC). Determination of CCK-like bioactivity and immunoreactivity after HPLC was performed in parallel by bioassay and radioimmunoassay, respectively CCK-58 was the most abundant molecular form in the porcine brain extract, followed by CCK-33, and CCK-8. Both assay systems measured similar relative concentrations of all different molecular forms of CCK. To elucidate the biological potency of CCK-58 before and after tryptic cleavage, a CCK-58-enriched fraction was prepared by HPLC. Part of this material was quantitatively cleaved by trypsin resulting in the formation of small molecular forms of CCK. Bioactivity and immunoreactivity of equal amounts of cleaved and uncleaved material (= CCK-58) were determined by bioassay and radioimmunoassay in parallel. Cleavage of CCK-58 increased CCK-like bioactivity by 260% and CCK-like immunoreactivity by 310%. These results indicate that in a rat pancreatic acini system, porcine CCK-58 exerts 25-30% bioactivity compared to smaller CCK forms. The specific CCK antiserum G-160 seems to possess 25-30% affinity to CCK-58 in comparison to trypsin-cleaved CCK-58. It can be concluded that this antiserum exhibits an affinity to different molecular forms of CCK which parallels their relative bioactivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗