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Biomedical subjects

P A Johnson

Publications and source records attributed to P A Johnson.

At least 19 recordsLinked to original sources
PubMed2006

Prophylactic internal fixation of the radial osteocutaneous donor site.

The technique of prophylactic internal fixation (PIF) of the radial osteocutaneous donor site is described and reviewed. Twenty-two donor sites were reinforced with a 3.5mm dynamic compression plate across the anterior defect. The incidence of fracture was 4.5% (1 out of 22). The single fracture was due to a technical error and was managed conservatively. Fracture at a donor site that has not been plated is more frequent and often becomes displaced, requiring secondary surgery. In contrast, the incidence of fracture, displacement and secondary surgery following prophylactic internal fixation (PIF) is relatively low. There have been no significant long-term complications with PIF. It is now the method of choice for managing the radial osteocutaneous donor site.

Bone Plates↗
PubMed2006

Bone micro-damage assessment using non-linear resonant ultrasound spectroscopy (NRUS) techniques: a feasibility study.

Non-linear resonant ultrasound spectroscopy (NRUS) is a technique exploiting the significant non-linear behavior of damaged materials, related to the presence of damage. This study shows for the first time the feasibility of this technique for damage assessment in bone. Two samples of bovine cortical bone were subjected to a progressive damage experiment. Damage accumulation was progressively induced in the samples by mechanical testing. For independent assessment of damage, X-ray CT imaging was performed at each damage step, but only helped in the detection of the prominent cracks. Synchrotron micro-CT imaging and histology using epifluorescence microscopy were performed in one of the two samples at the last damage step and allowed detection of micro-cracks for this step. As the quantity of damage accumulation increased, NRUS revealed a corresponding increase in the non-linear response. The measured change in non-linear response is much more sensitive than the change in elastic modulus. The results suggest that NRUS could be a potential tool for micro-damage assessment in bone. Further work has to be carried out for a better understanding of the physical nature of damaged bone, and for the ultimate goal of in vivo implementation of the technique where bone access will be a challenging problem.

Algorithms↗
PubMed2006

Cyclooxygenase 1 and 2 mRNA and protein expression in the Gallus domesticus model of ovarian cancer.

OBJECTIVE: Our purpose was to determine the mRNA and protein expression of cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2) in ovarian tumors and normal ovaries of the hen, which is an excellent model for human ovarian cancer. Tissue concentrations of prostaglandin E(2) (PGE2) and PGE2 metabolites were also determined. METHODS: Tissue was obtained from ovarian tumor (n = 18) and normal ovary (n = 29) of 2- to 4-year old Single-comb White Leghorn hens. Quantitative real-time PCR with Sybr Green was used to quantify the mRNA expression of COX-1 and COX-2, using 18S expression as an internal control for COX normalization. Immunohistochemistry using antibodies for COX-1 and COX-2 was used to localize protein expression of each isoform in a subset of tumor (n = 5) and normal samples (n = 6). For determination of tissue prostaglandin concentration, tissue was obtained from ovarian tumor (n = 8) and normal ovary (n = 8). PGE2 and PGE2 metabolites were measured using competitive enzyme immunoassays (EIAs). RESULTS: Our results indicate that COX-1 mRNA expression is significantly higher (P < 0.05) in ovarian tumor samples compared to normal ovaries while there is no significant difference in expression of COX-2 between the samples. Immunohistochemistry results support this finding and show COX-1 expression only in tumor samples and COX-2 expression unchanged between normal ovary and tumor samples. PGE2 levels are significantly higher (P < 0.05) in tumor samples compared to normal ovaries, and there is no significant difference in PGE2 metabolite levels between the samples. CONCLUSION: These findings may implicate COX-1 as a suitable target for the prevention or treatment of ovarian cancer.

Animals↗
PubMed2006

Effect of activin A and inhibin A on expression of the inhibin/activin beta-B-subunit and gonadotropin receptors in granulosa cells of the hen.

Activin A has been shown to be abundant in the theca layer of the large pre-ovulatory follicles of the hen whereas inhibin A is produced in the granulosa layer. The purpose of this study was to investigate the effects of activin A and inhibin A on granulosa cell expression of inhibin beta-B-subunit, FSH receptor (FSHR), and LH receptor (LHR). Granulosa cells were isolated from the F1, F3+F4, and small yellow follicles (SYF; 6-12 mm diameter) of laying hens and pooled according to size. The cells were dispersed and plated in the presence of 0, 10, or 50 ng/ml recombinant human activin A (n=5 replicate cultures). RNA was subsequently extracted from the cells and Northern blots performed. Cell proliferation was determined for all treatments. An identical set of experiments was performed in which the granulosa cells were treated with recombinant human inhibin A (n=4 replicate cultures). Treatment with activin A at 50 ng/ml significantly (p<0.05) increased expression of beta-B-subunit for granulosa cells from all follicles. This dose also significantly increased expression of FSHR in granulosa cells from all follicles (p<0.05) and increased expression of LHR in cells from F1 and F3+F4 follicles (p<0.01) with no significant effect on cells from the SYF. Overall, activin A treatment significantly (p<0.05) decreased cell proliferation at the 50 ng/ml dose. Inhibin A had no significant effect on expression of beta-B-subunit, FSHR or LHR at any dose. There was a moderate stimulatory effect of inhibin A on granulosa cell proliferation. These results suggest that activin A may have an important role in regulating granulosa cell responsiveness to gonadotropins while also modulating follicle development by attenuating cell proliferation.

Activins↗
PubMed2006

Use of genetic strains of chickens in studies of ovarian cancer.

Ovarian cancer is a deadly disease often diagnosed late in development when there is little chance for a successful recovery. Although ovarian cancer is a rare occurrence in most animals, the domestic hen has been shown to spontaneously develop the disease with an age-related incidence. Two strains of hens derived from a similar genetic background and maintained at Cornell University have been shown to differ in the incidence of the disease. At 2 yr of age, the C strain hens have a greater incidence of ovarian neoplasms than do K strain hens. Interestingly, levels of plasma estradiol are elevated in the C strain compared with K strain hens. In addition, plasma immunoreactive inhibin is lower in the C strain than in the K strain. Finally, mRNA expression of the alpha-subunit of inhibin in the granulosa cell layer of the large yellow follicles is lower in the C strain compared with the K strain hens. Further studies using these as well as other strains of hens may be useful in learning more about the etiology of this disease.

Animals↗
PubMed2005

Pattern of secretion of immunoreactive inhibin/activin subunits by avian granulosa cells.

The messenger RNA expression for the inhibin/activin subunits in the granulosa layer of avian follicles of different developmental stages has previously been reported. In the present study, we examined the pattern of secretion of these protein subunits from cultured granulosa cells (GC) of avian follicles of defined maturity. Laying hens were euthanized and the F1, F2, F3, F4, small yellow follicles (SYF; 6-10 mm) and large white follicles (LWF; 3-5 mm) were removed. GC were isolated from the follicles, plated by size at a density of 6.25 x 10(5)cells per well (3 wells per follicle size) and cultured for 48 h in medium 199 with 5% FBS, antibiotics, and 1.0 microg/ml bovine insulin. After 48 h, the cultures were terminated and the media were saved (n = 6 replications). Proteins were precipitated from media, reconstituted for electrophoresis (SDS-PAGE), and analyzed by Western blot. Progesterone was also measured in the medium. For detection of the inhibin alpha-subunit, a rabbit antibody against the chicken inhibin alpha-subunit (1-26 aa) was used. The betaA-subunit was detected with rabbit anti-betaA-subunit (81-113 aa) and the betaB-subunit was detected with rabbit anti-betaB-subunit (80-112 aa). Under reduced conditions, GC from the larger follicle sizes (F1-F4) secreted the most (p < 0.05) immunoreactive inhibin alpha-subunit compared to smaller follicle sizes. Under non-reduced conditions, a band at approximately 32 kDa was detected by both the alpha-subunit antibody and by the betaA-subunit antibody in media from GC of the F1-F4 follicles, suggesting secretion of intact inhibin A. Immunoreactive alpha-subunit and betaB-subunit were detected under reduced conditions in media from the GC of the SYF, suggesting that this follicle population may secrete intact inhibin B. In addition, under non-reduced conditions, cells from the SYF secreted the greatest amount of intact inhibin B (p < 0.05) at a size of approximately 32 kDa. Cells from the LWF expressed low levels of all inhibin subunits. Progesterone concentration in the media from the F1 follicle was greatest and was decreased in media from smaller follicles. It is suggested that the largest follicles in the hierarchy are the source of most circulating intact inhibin A while the small follicles are the source of intact inhibin B.

Activins↗
PubMed2005

Does an oral analgesic protocol improve pain control for patients with cancer? An intergroup study coordinated by the Eastern Cooperative Oncology Group.

BACKGROUND: Cancer pain is highly prevalent and commonly undertreated. This study was designed to determine whether dissemination of a clinical protocol for pain management would improve outcomes in community oncology practices. PATIENTS AND METHODS: A pain management protocol was developed based on accepted guidelines. After baseline assessment, oncology practices were randomly assigned to 'analgesic protocol' (AP) sites, where oncologists implemented the guidelines in a group of lung or prostate cancer patients, or to 'physician discretion' (PD) sites, where customary treatment was continued. Patients treated on protocol and a comparison group of patients with pain due to breast cancer or myeloma were monitored for change in pain using the Brief Pain Inventory, and for change in other symptoms or mood. RESULTS: The protocol terminated early because of poor accrual. We compared groups using proportions of patients who had no or mild pain at follow-up. Although measures of protocol adherence did not suggest the occurrence of major practice change, the proportion of lung or prostate cancer patients with no or mild pain increased significantly from baseline for those treated at AP sites compared with those treated at PD sites. There was no significant difference between the breast and myeloma patients treated at AP sites versus those treated at PD sites. CONCLUSION: A protocol for cancer pain management can improve pain control. Diffusion of these benefits to other patients was not confirmed. Given the small sample size, these findings require confirmation in a larger trial.

Administration, Oral↗
PubMed2004

Expression and function of growth differentiation factor-9 in an oviparous species, Gallus domesticus.

Many studies have indicated a critical role for the oocyte growth factor, growth differentiation factor-9 (GDF9), in mammalian follicle development, but no information has been available concerning oviparous species. We cloned a cDNA for chicken GDF9 (162 base pairs) and used it in Northern blot analysis to identify a transcript at 1.7 kilobase in RNA isolated from the ovary of the hen. We also sequenced two full-length clones from a normalized chicken reproductive tract cDNA library. The cDNA clone for chicken GDF9 encodes a protein of approximately 449 amino acids and all six cysteine residues, and three of the four glycosylation sites are conserved with respect to mammalian GDF9. Chicken GDF9 is approximately 65% similar in the full-length cDNA sequence and 80% similar in amino acid sequence at the C-terminal region to GDF9 from several mammals. Quantitative polymerase chain reaction analysis (n = 5) indicated that GDF9 mRNA is greatest in follicles < 1 mm in size compared with larger follicles or granulosa layers isolated from larger follicles. Immunocytochemical analysis showed strong expression of GDF9 in hen oocytes. In yolk-filled oocytes, the GDF9 was localized at the periphery of the oocyte. Finally, oocyte-conditioned medium (from < 1-mm oocytes) resulted in a 2-fold increase in granulosa cell proliferation, which could be inhibited by preincubation of the conditioned medium with GDF9 antibody. These data suggest that GDF9 is present in the hen oocyte and that this factor is capable of enhancing granulosa cell proliferation, as has been demonstrated in mammals.

Amino Acid Sequence↗
PubMed2002

Apoptosis and proliferation of acinar and islet cells in chronic pancreatitis: evidence for differential cell loss mediating preservation of islet function.

BACKGROUND: Chronic pancreatitis is characterised clinically by early exocrine insufficiency, with diabetes mellitus occurring as a late phenomenon. This is mirrored pathologically by extensive acinar cell destruction and islet preservation. The mechanisms underlying this differential rate of cellular destruction are unknown. AIMS: To test the hypothesis that acinar loss and islet preservation in chronic pancreatitis occurs due to differential epithelial kinetics and investigate the role of inflammatory cells and cell cycle associated molecules. METHODS: Archival tissue from six chronic pancreatitis cases was compared with six normal controls using TUNEL and immunohistochemistry for CD3, CD20, CD68, MIB-1, Bcl-2, Bax, Fas, Fas ligand, retinoblastoma protein (Rb), and tissue inhibitor of metalloproteinases 1 (TIMP-1) and 2 (TIMP-2). RESULTS: The acinar cell apoptotic index (AI) and proliferation index were higher in chronic pancreatitis than controls. T lymphocytes diffusely infiltrated fibrous bands and acini but rarely islets. Acinar Bcl-2 expression exceeded islet expression in chronic pancreatitis and controls while Bax was strongly expressed by a subset of islet cells and weakly by centroacinar cells. Islet Fas and Fas ligand expression exceeded acinar expression in chronic pancreatitis and controls. Acinar Rb expression was higher in chronic pancreatitis than in controls. Islets in chronic pancreatitis and controls showed intense TIMP-1 and TIMP-2 expression. CONCLUSION: Apoptosis plays a significant role in acinar loss in chronic pancreatitis. Acinar Bcl-2 and islet Bax expression indicates complex AI control. Increased acinar Rb expression in chronic pancreatitis may differentially promote acinar loss. Fas ligand expression may be restricted to islet cell membranes through TIMP-1 expression and inhibit islet damage by promoting apoptosis of cytotoxic T lymphocytes.

Apoptosis↗
PubMed2001

An in vivo/in vitro comparison with a leuprolide osmotic implant for the treatment of prostate cancer.

An osmotically driven implantable system was designed and characterized for the delivery of leuprolide over a year-long duration. Leuprolide has been used in the treatment of prostate cancer since the 1980s. The DUROS implant consists of a titanium alloy cylinder, measures 4 mm in diameter by 45 mm in length and holds approximately 150 microl of formulation. Stability studies indicated that leuprolide was stable, as a solution formulation in DMSO, for several years at 37 degrees C. In vitro release rate testing, at weekly intervals, showed zero-order delivery for 1 year. DUROS implants demonstrated release rates that were reproducible and similar to one another after storage at 25 degrees C for 18 months prior to testing. In vivo studies, with implants placed subcutaneously, revealed delivery rates comparable to those observed under in vitro conditions. Leuprolide stability was also comparable between in vivo and in vitro conditions. Steady leuprolide serum levels produced by the implant resulted in the desired pharmacodynamic efficacy endpoint of testosterone suppression, both in canines and in humans. The good agreement between in vivo/in vitro delivery rates was as expected for a delivery system based on the principles of osmosis.

Alloys↗
PubMed2001

Oncolytic herpes simplex virus vector with enhanced MHC class I presentation and tumor cell killing.

Oncolytic herpes simplex virus type 1 (HSV-1) vectors are promising therapeutic agents for cancer. Their efficacy depends on the extent of both intratumoral viral replication and induction of a host antitumor immune response. To enhance these properties while employing ample safeguards, two conditionally replicating HSV-1 vectors, termed G47Delta and R47Delta, have been constructed by deleting the alpha47 gene and the promoter region of US11 from gamma34.5-deficient HSV-1 vectors, G207 and R3616, respectively. Because the alpha47 gene product is responsible for inhibiting the transporter associated with antigen presentation (TAP), its absence led to increased MHC class I expression in infected human cells. Moreover, some G47Delta-infected human melanoma cells exhibited enhanced stimulation of matched antitumor T cell activity. The deletion also places the late US11 gene under control of the immediate-early alpha47 promoter, which suppresses the reduced growth properties of gamma34.5-deficient mutants. G47Delta and R47Delta showed enhanced viral growth in a variety of cell lines, leading to higher virus yields and enhanced cytopathic effect in tumor cells. G47Delta was significantly more efficacious in vivo than its parent G207 at inhibiting tumor growth in both immune-competent and immune-deficient animal models. Yet, when inoculated into the brains of HSV-1-sensitive A/J mice at 2 x 10(6) plaque forming units, G47Delta was as safe as G207. These results suggest that G47Delta may have enhanced antitumor activity in humans.

Animals↗
PubMed2001

Cytokine gene transfer enhances herpes oncolytic therapy in murine squamous cell carcinoma.

Replication-competent, attenuated herpes simplex viruses (HSV) have been demonstrated to be effective oncolytic agents in a variety of malignant tumors. Cytokine gene transfer has also been used as immunomodulatory therapy for cancer. To test the utility of combining these two approaches, two oncolytic HSV vectors (NV1034 and NV1042) were designed to express the murine GM-CSF and murine IL-12 genes, respectively. These cytokine-carrying variants were compared with the analogous non-cytokine-carrying control virus (NV1023) in the treatment of murine SCC VII squamous cell carcinoma. All three viruses demonstrated similar infection efficiency, viral replication, and cytotoxicity in vitro. SCC VII cells infected by NV1034 and NV1042 effectively produced GM-CSF and IL-12, respectively. In an SCC VII subcutaneous flank tumor model in immunocompetent C3H/HeJ mice, intratumoral injection with each virus caused a significant reduction in tumor volume compared with saline injections. The NV1042-treated tumors showed a striking reduction in tumor volume compared with the NV1023- and NV1034-treated tumors. On subsequent rechallenge in the contralateral flank with SCC VII cells, 57% of animals treated with NV1042 failed to develop tumors, in comparison with 14% of animals treated with NV1023 or NV1034, and 0% of naive animals. The increased antitumor efficacy seen with NV1042 in comparison with NV1023 and NV1034 was abrogated by CD4(+) and CD8(+) lymphocyte depletion. NV1042 is a novel, attenuated, oncolytic herpesvirus that effectively expresses IL-12 and elicits a T lymphocyte-mediated antitumor immune response against murine squamous cell carcinoma. Such combined oncolytic and immunomodulatory strategies hold promise in the treatment of cancer.

Animals↗
PubMed2001

Cephalosporins can be given to penicillin-allergic patients who do not exhibit an anaphylactic response.

STUDY OBJECTIVES: To assess the risk of intraoperative allergic reactions to cephalosporins in patients who claim to be allergic to penicillin. DESIGN: Retrospective chart review. SETTING: University-affiliated hospital. MEASUREMENTS: 2,933 intraoperative anesthesia records of all adult orthopedic patients treated at our institution during a 14-month period (7/96-8/97) were reviewed for antibiotic use and allergic reactions. MAIN RESULTS: Most of the 2,933 orthopedic patients, including 413 patients who were allergic to penicillin, received a cephalosporin (usually cefazolin) during their procedure. Only one of the penicillin-allergic patients may have had an allergic reaction to the cephalosporin, because diphenhydramine and hydrocortisone were given at the beginning of the case. However, no mention was made on the chart about itching or a rash or hives. One of the non-penicillin-allergic patients did develop a rash while the cephalosporin was infusing, requiring discontinuation of the antibiotic. CONCLUSIONS: Given the low incidence of allergic reactions, it appears to be safe to administer cephalosporins to patients who claim to be allergic to penicillin. However, no conclusion can be made concerning patients who report severe or anaphylactic reactions to penicillin, because these patients probably were excluded from the study.

Anaphylaxis↗
PubMed2001

Preferences for quality of life or survival expressed by patients with heart failure.

BACKGROUND: As many patients with heart failure develop symptoms limiting daily life, newer therapies may be found to improve functional status without concomitant survival benefit. As some of these therapies may actually increase mortality, it is increasingly relevant to assess patients' preferences for survival vs improvement in symptoms. METHODS: We enrolled 99 patients with advanced heart failure (ejection fraction 24 +/- 10, duration 6 +/- 5 years). Each patient completed time trade-off and standard gamble instruments, Minnesota Living with Heart Failure questionnaires and visual analog scales for dyspnea and overall health. Jugular venous pressure was assessed in all patients and peak oxygen consumption was measured during bicycle exercise in 60 patients. RESULTS: Strong polarity of preference toward either survival or quality of life was expressed by 60% of patients. There was good correlation between time trade-off and standard gamble utility scores (r = 0.64), and between preference and functional class (r = 0.60). Higher jugular venous pressure and lower peak oxygen consumption were associated with poorer utility scores (p <.05). Higher dyspnea scores and worse Living with Heart Failure scores were also associated with preference to trade time or take risks for better health. CONCLUSIONS: These findings suggest that heart failure patients express meaningful preferences about quality vs length of life. High jugular venous pressure, low peak oxygen consumption and poor Living with Heart Failure scores were related to low utility scores. These cannot be assumed, however, to predict the intensity of individual preference to trade nothing or virtually everything for better health.

Adult↗
PubMed2001

Effective treatment of head and neck squamous cell carcinoma by an oncolytic herpes simplex virus.

BACKGROUND: The prognosis of patients with advanced or recurrent head and neck squamous cell carcinoma remains poor despite refinements in multimodality therapies. This study evaluates the efficacy of a replication-competent, attenuated, oncolytic herpes simplex virus, NV1020, as a novel agent in the treatment of human head and neck squamous cell carcinoma (HNSCC). STUDY DESIGN: Five different HNSCC lines were exposed to NV1020 in vitro at varying viral concentrations. The ability of the virus to lyse and replicate within these cancer cells in vitro was determined by cytotoxicity assay and plaque assay, respectively. Three HNSCC lines were grown in the subcutaneous flanks of athymic nude mice and treated with an intratumoral injection of NV1020 or saline as a control. Tumor dimensions were subsequently measured at serial time points and tumor volumes were calculated. Herpes simplex virus (HSV)-1 immunohistochemistry was performed on excised tumors to determine the efficacy of in vivo tumor infection by NV1020. RESULTS: NV1020 was highly cytotoxic in vitro to all five human HNSCC lines at a concentration of one infectious viral particle per cancer cell, and had variable cytotoxicity at a 100-fold lower concentration. Viral replication in vitro by NV1020 was efficient in four of five HNSCC lines with a greater than 200-fold increase in viral titers. Flank tumors treated with intratumoral injections of NV1020 resulted in significant regression of all tested HNSCC lines. HSV-1 immunohistochemistry of excised flank tumors treated with NV1020 demonstrated positive cytoplasmic staining and areas of tumor necrosis at 24 hours after injection. CONCLUSIONS: NV1020 is an oncolytic HSV that displays efficient replication and oncolysis in human HNSCC lines in vitro. Injection of NV1020 into murine flank tumors demonstrated effective tumor regression. Treatment of HNSCC with NV1020 is a promising form of therapy with potential clinical applicability in humans.

Animals↗
PubMed2001

Follicle-stimulating hormone regulation of inhibin alpha- and beta(B)-subunit and follistatin messenger ribonucleic acid in cultured avian granulosa cells.

FSH regulation of inhibin alpha-, beta(B)-subunit and follistatin mRNA was investigated in cultured chicken granulosa cells, which were isolated and pooled according to size from the F(4) + F(5) follicles, small yellow follicles (SYF), and large white follicles (LWF). In experiment 1 (four replicate experiments), granulosa cells were cultured, and the effect of FSH (50 ng/ml) on the growth of cells from the different follicles was examined at 24 and 48 h of culture. Cell viability was >95% for all of the granulosa cell cultures at 24 and 48 h. At 24 h, the number of granulosa cells in both the FSH-treated and the untreated cultures for all follicle types was numerically greater than the number of cells originally plated. At 48 h, FSH-treated cultures for all follicle types had twice (P: < 0. 05) the number of cells as the untreated cultures. In experiment 2 (three replicate experiments), FSH increased expression of the mRNA for inhibin alpha-subunit in LWF granulosa cells at 4 and 24 h to detectable levels and increased inhibin alpha-subunit protein accumulation to detectable levels by 24 h in granulosa cells from the LWF. FSH also increased (P: < 0.05) mRNA levels for the inhibin alpha-subunit at 4 and 24 h in SYF granulosa cells and at 24 h in F(4) + F(5) granulosa cells. The effects of FSH on follistatin and ss(B)-subunit were variable with respect to follicle development and culture duration. These results suggest that FSH plays an important role in stimulating the production of mRNA and protein for the inhibin alpha-subunit in small prehierarchical follicles.

Animals↗
PubMed2001

Activin A and gonadotropin regulation of follicle-stimulating hormone and luteinizing hormone receptor messenger RNA in avian granulosa cells.

Activin A regulation of the expression of mRNA for the LH receptor, FSH receptor, and the inhibin alpha subunit as well as the effect of activin A on the secretion of progesterone were investigated in chicken granulosa cell cultures. Granulosa layers were isolated from the F(1) and F(3) + F(4) follicles from five hens, pooled according to size, dispersed, and cultured for 48 h. In experiment 1 (n = 3 replications), granulosa cells were cultured with or without highly purified ovine (o) FSH at 50 ng/ml and in the presence of 0, 10, or 50 ng/ml of recombinant chicken activin A. Experiment 2 (n = 4 replications) followed the same protocol as experiment 1, except that oFSH was replaced with oLH. Results from these experiments showed that addition of activin A to the granulosa cell cultures had no effect on the expression of mRNA for the inhibin alpha subunit or the FSH receptor, but it did affect the expression of mRNA for the LH receptor. Treatment of F(3) + F(4) granulosa cells with LH stimulated the expression of mRNA for the LH receptor; however, when LH was combined with either dose of activin A, this induction was prevented. The highest dose of activin A with or without LH resulted in decreased expression of the LH receptor compared to the untreated controls in the F(3) + F(4) cell cultures. Progesterone secretion by the granulosa cells from both follicle sizes was not altered by activin A. In experiment 3 (n = 3 replications), the effect of activin A on the growth of granulosa cells was examined with the following treatments: 0, 10, or 50 ng/ml of activin A; 50 ng/ml of either oLH or oFSH; and oLH or oFSH combined with 10 ng/ml of activin A. The highest dose of activin reduced the rate of granulosa cell proliferation in both follicle types. Growth of F(1) and F(3) + F(4) granulosa cells was stimulated by the addition of either gonadotropin, and the presence of 10 ng/ml of activin A with either gonadotropin did not alter this proliferation, except for the LH-treated F(3) + F(4) granulosa cells, in which the increase in proliferation was prevented. The results suggest that activin A could act as a local factor that regulates follicular maturation by preventing excessive or untimely LH receptor expression.

Activins↗