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J M Hammond

Publications and source records attributed to J M Hammond.

At least 55 records · Page 3Linked to original sources
PubMed1994

IGF-binding proteins are differentially regulated in an ovarian granulosa cell line.

We have recently established an immortalized granulosa cell line as a model system to investigate ovarian function, with particular emphasis on the insulin-like growth factor (IGF) regulatory system. Previous results have shown that these cells express mRNAs for IGF-binding proteins (IGFBPs)-2 to -5. These IGFBPs are also detected by ligand blots. The current work evaluated the regulation by the IGFs and cAMP on the IGFBPs and their mRNAs and compared the findings to that in primary culture. Our results indicate that levels of the IGFBPs are controlled, in part, by expression of the mRNAs. However, evidence for post-transcriptional regulation was also discovered. IGFBP-3 was stimulated by IGF-I, IGFBP-4 by forskolin, and IGFBP-5 by IGF-I. IGFBP-2, -3, and -4 are expressed under basal conditions whereas IGFBP-5 is only detectable after IGF-I induction. An alteration in the biphasic actions of cAMP in this cell line, as compared to primary culture, was evident.

Animals↗
PubMed1994

Microbiological surveillance during selective decontamination of the digestive tract (SDD)

The use of selective decontamination of the digestive tract (SDD) as prophylaxis against nosocomial respiratory tract infection remains controversial, largely because of concerns that, in the long term, it may promote the emergence of antibiotic-resistant strains. This report describes the results of surveillance cultures and susceptibility testing undertaken during the course of a 2-year, double-blind study of the efficacy of SDD which was conducted in a respiratory intensive care unit (ICU). Surveillance specimens from the alimentary tract and trachea were obtained from each patient on admission and then twice weekly until 48 h after discharge from the unit. Specimens were cultured semiquantitatively and organisms from morphologically distinct colonies were identified by standard methods; the susceptibilities of these isolates were determined by the disc diffusion method. Five thousand, nine hundred and sixty surveillance samples from 239 patients were processed in this way. Compared with the placebo group, SDD caused a significant reduction in the incidence of colonization of the alimentary tract with aerobic Gram-negative bacilli (AGNB), and Candida spp. were almost totally eliminated. The incidence of colonization with enterococci increased in both groups, while the incidence of both colonization of the alimentary tract with strains of coagulase-negative staphylococci and methicillin-resistant Staphylococcus aureus and infection caused by these organisms increased in the SDD group. Acinetobacter spp. were the most common bacteria associated with unit-acquired colonization and lower respiratory tract infection in both groups. The acquisition of strains of Pseudomonas aeruginosa and cefotaxime- and/or tobramycin-resistant Enterobacteriaceae was significantly greater in the placebo group than in the SDD group, although tobramycin-resistant strains of Proteus, Morganella and Providencia spp. were isolated from three of 114 patients receiving SDD. The use of SDD did not lead to an overall increase in antibiotic resistance amongst the AGNB usually associated with ICU-acquired infection. However, colonization with strains which were either resistant to one or more of the antibiotic components of the regimen or which were not inhibited by the regimen was observed and may subsequently lead to infection.

Anti-Bacterial Agents↗
PubMed1994

Effects of growth hormone and gonadotropin on the insulin-like growth factor system in the porcine ovary.

The effects of growth hormone (GH) +/- pregnant mare's serum gonadotropin (PMSG) on levels of insulin-like growth factor (IGF)-I and -II and IGF binding protein (BP)-2 and -3 in serum and follicular fluid (FFI) and on the expression of their mRNA in the ovaries of prepubertal gilts were determined. Steroids in FFI were also quantified. In the first experiment, GH, given for either 20 or 40 days, caused a distinct (threefold, p < 0.05) increase in IGF-I in both serum and FFI with no change in the FFI:serum ratio (0.65). Effects of GH on IGF-II were opposite, with a drop in circulating and FFI levels (p < 0.05). In contrast to data for IGF-I, FFI levels were higher than those in serum for IGF-II (1.42, FFI:serum); IGF-II levels and the ratio fell after GH treatment. GH for either 20 days or 40 days increased serum IGBP-3 to 140% and 250% of control values while decreasing serum IGFBP-2 by 46% and 31%, respectively (p < 0.001). FFI IGFBP-3 was increased to a similar extent by GH (p < 0.005), but IGFBP-2 was not affected. Neither progesterone (P4) nor estradiol (E2) was affected by treatment with GH. However, androstenedione (A4) was decreased by 20-day and 40-day GH treatment relative to the respective controls (p < 0.05). In the second experiment, PMSG resulted in a modest (28%) increase in intrafollicular IGF-I (p < 0.06).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Porcine ovarian granulosa cells secrete insulin-like growth factor-binding proteins-4 and -5 and express their messenger ribonucleic acids: regulation by follicle-stimulating hormone and insulin-like growth factor-1.

Using ligand blotting, Western immunoblotting, and Northern analysis, we have characterized the insulin-like growth factor (IGF)-binding proteins (IGFBPs) produced by cultures of porcine granulosa cells. Ligand blot analysis of conditioned medium from untreated cultures of moderately differentiated granulosa cells (MDGCs; from 4-6-mm follicles) revealed mainly IGF-binding activity associated with IGFBP-2 (34 kDa) and IGFBP-3 (40/44-kDa doublet), which have previously been identified and characterized. In addition, these cultures secreted 30- and 22-kDa forms under some circumstances. The identification and regulation of these IGFBPs of lower molecular mass were the focus of the current studies. Treatment of these MDGCs with IGF-I dramatically stimulated the production (to a detectable level) of a 30-kDa IGFBP that was identified by immunoblotting with antiserum to IGFBP-5 but not antisera to IGFBP-1, -2, -3, -4, or -6. Production of IGFBP-5 was attenuated by concurrent treatment with FSH. IGFBP-5 mRNA in these cultures was correspondingly stimulated by IGF-I but unaffected by FSH. FSH increased the level of a minor 22-kDa IGFBP. Messenger RNAs for IGFBP-1, -4, and -6 were also examined but only IGFBP-4 mRNA was detectable, suggesting that the 22-kDa band was IGFBP-4. These results were compared to those in cultures of immature granulosa cells from 1-3-mm follicles, in which 22- and 30-kDa IGFBPs were readily detectable. An antiserum to IGFBP-4 precipitated the 22- and 30-kDa bands whereas deglycosylation shifted the 30-kDa IGFBP to 22 kDa, suggesting that both these bands represent glycosylation variants of IGFBP-4.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Changes in the messenger ribonucleic acid for insulin-like growth factor-I and -II in the porcine testis during and between two waves of testicular development.

The boar testis was used as a model for examining the possible role of production of the insulin-like growth factors (IGF) in steroidogenesis and/or testicular growth as testicular development occurs in waves. Blood and testes were sampled from boars at different ages (100-102 days of gestation; 7, 19, and 30 days; and 10 and 25 wk), selected to occur during and between the last two waves of testicular development. Serum was analyzed for testosterone and RNA was extracted from the testes for Northern and dot-blot analysis of IGF mRNA. Testosterone concentrations declined (p = 0.01) from 7 days to 10 wk of age and rebounded at 25 wk, indicating completion of the second and third waves of testicular development. The quantity of testicular mRNA for IGF-I increased gradually with age, and that for IGF-II decreased. We therefore conclude that regulation of the expression of the mRNAs for IGF-I and -II in the pig testis is not a function of either the waves of testicular development or the level of steroidogenesis.

Actins↗
PubMed1994

Steroidogenic cytochrome P450 enzyme messenger ribonucleic acids and follicular fluid steroids in individual follicles during preovulatory maturation in the pig.

Changes in follicular concentrations of steroidogenic cytochrome P450 enzyme mRNAs were determined during preovulatory maturation. RNA was isolated from 59 individual follicles dissected from 18 pigs during altrenogest-synchronized preovulatory follicular maturation: at Day 1 (pre-follicular phase), Day 3 (early follicular phase), Day 5 (mid-follicular phase), and Day 7 (late follicular phase, 24-36 h after the onset of the LH surge). Follicular fluid was aspirated for steroid RIA. RNA was also isolated from pooled granulosa cells, theca tissue, and luteal tissue. RNA was analyzed by Northern and slot-blot procedures using cDNA probes to human aromatase cytochrome P450 (P450arom), porcine 17 alpha-hydroxylase cytochrome P450 (P450(17) alpha), and porcine cholesterol side-chain cleavage cytochrome P450 (P450scc). P450arom mRNA was expressed in both granulosa and theca interna cells but was not detectable in luteal tissue from Day 13 of the estrous cycle. Follicular P450arom mRNA concentration tended to increase between Days 1 and 5 and decreased (p < or = 0.05) by 92% between Days 5 and 7. Follicular fluid estradiol-17 beta concentration increased 17-fold between Days 1 and 5 and then decreased (p < or = 0.05) by 96% between Days 5 and 7. P450(17) alpha mRNA was present in theca interna but was not detected in granulosa cells or luteal tissue. Follicular P450(17) alpha mRNA concentrations did not differ significantly among days, but the content per follicle increased (p < 0.05) between Days 1 and 5 and decreased (p < 0.05) between Days 5 and 7.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Selective decontamination of the digestive tract in multiple trauma patients--is there a role? Results of a prospective, double-blind, randomized trial.

OBJECTIVE: To evaluate the efficacy of the technique of selective decontamination of the digestive tract in preventing the development of secondary infection and its influence on morbidity and mortality rates in multiple trauma patients with chest injuries requiring intermittent positive-pressure ventilation. DESIGN: Prospective, double-blind, randomized study. SETTING: A multidisciplinary respiratory intensive care unit (ICU) in a 1,500-bed teaching hospital. PATIENTS: Seventy-two patients (mean Injury Severity Score of 29.5) who were intubated for > 48 hrs and remained in the ICU for > 5 days. INTERVENTIONS: Patients were randomized on admission to receive selective decontamination therapy or placebo. All patients received intravenous cefotaxime for 72 hrs and the treatment group received oral and enteral selective decontamination with amphotericin B, polymyxin E, and tobramycin (n = 39), while the placebo group received a placebo containing oral paste and enteral solution (n = 33). MEASUREMENTS: Secondary infection was determined clinically and microbiologically and surveillance cultures were monitored for gastrointestinal colonization. RESULTS: The patient groups were fully comparable for age, severity of illness, and compromising factors. There was no difference in the number of patients infected (11 treatment group vs. 11 placebo), infections (17 vs. 16) and deaths (5 vs. 3); the duration of ICU (15.5 vs. 14.2 days) and hospital stays (26.3 vs. 25.5) were also similar. Microbiological surveillance cultures confirmed effective elimination of aerobic Gram-negative bacilli, and infections in the treatment group were largely due to Staphylococcus aureus and Staphylococcus epidermidis. CONCLUSION: We have been unable to show any benefit from the use of selective decontamination of the digestive tract in the prevention of secondary infections in multiple trauma patients.

Adult↗
PubMed1994

Nucleotide sequence of a 55 kbp region from the right end of the genome of a pathogenic African swine fever virus isolate (Malawi LIL20/1).

The nucleotide sequence of a 55098 bp region from the right end of the genome of a virulent African swine fever virus (ASFV) isolate (Malawi LIL20/1) has been determined. Translation of the sequence identified 67 major open reading frames (ORFs) which are closely spaced and read from both DNA strands. At six positions intergenic tandem repeat arrays are found. Comparison of the predicted amino acid sequences of encoded proteins with protein sequence databases identified a number of homologies. These include three subunits of RNA polymerase, a protein with homology to transcription factor SII (TFSII), a DNA ligase, two subunits of mRNA capping enzyme, a DNA topoisomerase type II, a dUTPase, a protein kinase, three helicases, a ubiquitin-conjugating enzyme, a protein with homology to the nif S and nif S-like proteins identified in some bacteria and Saccharomyces cerevisiae, a protein with homology to both a myeloid differentiation primary response antigen (MyD116) and to a herpes simplex virus-encoded neurovirulence-associated protein (ICP34.5), a protein with homology to the ASFV-encoded structural protein p22, two proteins with homology to copies of the ASFV-encoded multigene family 360 and one protein with homology to the ASFV-encoded multigene family 110. Four genes encode proteins which have homology to each other and constitute a new multigene family (MGF100). Nine ORFs encode proteins which contain predicted transmembrane domains. The possible functions of these predicted ASFV-encoded proteins are discussed and the evolutionary relationship of ASFV to other viruses are considered. Despite the similarities in genome structure and replication strategy of ASFV with poxviruses, sequence similarity between them is low and the organization of ASFV-encoded genes is not colinear with that of the orthopoxviruses.

African Swine Fever Virus↗
PubMed1994

Proteolytic degradation of insulin-like growth factor (IGF)-binding protein-3 by porcine ovarian granulosa cells in culture: regulation by IGF-I.

Porcine ovarian granulosa cells in culture secrete glycosylated insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3), which inhibits gonadotropin and IGF action in the ovary. Synthesis of IGFBP-3 is stimulated by IGF-I and attenuated by gonadotropin. The purpose of the present study was to determine whether IGFBP-3 levels were also regulated via proteolysis. Exogenously added nonglycosylated recombinant human IGFBP-3 (rhIGFBP-3) was significantly degraded over time by a soluble serine-specific protease, similar to plasmin, in control cultures and those treated with FSH, insulin, or several other classes of hormones. In contrast, degradation was greatly attenuated by the IGFs. Degraded rhIGFBP-3 exhibited much reduced affinity for [125I]IGF-II, suggesting that degradation could make available IGFs for cellular interaction. The mechanism of IGFBP-3 protease inhibition by IGFs is unclear. Mediation by IGF receptors is unlikely, as insulin at a dose that activated both insulin and type I IGF receptors did not alter intrinsic degradation of IGFBP-3 (as does IGF). Additionally, IGF-I attenuation of IGFBP-3 degradation was not inhibited by antagonism of receptor action with a tyrosine kinase inhibitor. Further, IGF-I inhibited degradation in cell-free conditioned medium. Direct stabilization of IGFBP-3 via binding of IGFs was suggested from these results. However, long R3 IGF-I attenuated IGFBP-3 degradation even though it has low affinity for IGFBPs. Inhibition of the protease by IGFs is also possible. We conclude that IGFs inhibit the degradation of exogenous nonglycosylated rhIGFBP-3. If active in vivo, this may serve to increase endogenous IGFBP-3 levels in follicular fluid.

Animals↗
PubMed1994

Insulin-like growth factor-binding protein-2 and -3 are correlated with atresia and preovulatory maturation in the porcine ovary.

We compared insulin-like growth factor-binding protein (IGFBP) levels with indicators of follicular maturation and atresia in individual follicles of the porcine ovary. Follicular development was synchronized with the progestin, altrenogest, and progestin withdrawal was used to initiate the growth of an ovulatory cohort of follicles, which is accompanied by atresia of noncohort follicles. Individual follicles were isolated on days 1, 3, 5, and 7 after progestin withdrawal. Atretic follicles were identified by the presence of low hypodiploid levels of DNA in 10% or more of their granulosa cells using flow cytometry. The follicular fluid (FF) level of IGFBP-3 did not differ significantly between healthy and atretic medium-sized (3- to 6-mm) follicles and was not significantly correlated with the percentage of granulosa cells containing hypodiploid levels of DNA (r = 0.181) or with endocrine parameters such as FF concentrations of estradiol or androstenedione. However, among healthy follicles (atretic follicles removed from analyses to better examine follicular maturation), IGFBP-3 increased (P < 0.01) between days 1 and 7 and was positively correlated with follicle diameter (r = 0.514; P < 0.05) and the FF concentration of progesterone (r = 0.556; P < 0.01), indicators of the degree of follicular maturation. FF IGFBP-2 levels were 3-fold greater (P < 0.01) in atretic than in healthy follicles, and IGFBP-2 was correlated with percentage of granulosa cells containing hypodiploid levels of DNA (r = 0.729; P < 0.001). Among healthy follicles, FF IGFBP-2 did not differ significantly among days and was not significantly correlated with follicle diameter. These data suggest that the content of IGFBP-2 is related to the state of follicular health/atresia, whereas IGFBP-3 is related to preovulatory follicular development.

Animals↗
PubMed1993

African swine fever virus genome content and variability.

A 55 kilobase pair (kb) region from the right end of the virulent African swine fever virus isolate, Malawi LIL20/1, has been sequenced. The 68 major open reading frames (ORFs) encoded are generally closely spaced and read from both DNA strands across the complete sequence. Comparison of the amino acid sequences of predicted ORFs with sequence databases identified 15 ORFs which encode proteins that are similar to proteins of known function. Two ORFs are homologous to copies of multigene family 360 (MGF360) and one ORF is homologous to copies of multigene family 110 (MGF110). Both of these multigene families have been described previously.

African Swine Fever Virus↗
PubMed1993

Expression of the IGF system in primary and immortalized porcine ovarian granulosa cells.

We have established a novel granulosa cell line derived from porcine ovarian follicles (4-6 mm in diameter). This cell line, MDG2.1, was obtained by transfection of freshly cultured cells with the plasmid pSV3neo. Doubling time for MDG2.1 cells is 24-36 h. Northern analysis for RNAs of the insulin-like growth factor (IGF) regulatory system indicates that RNAs for IGF-I, IGF-I receptor, and IGF binding proteins (IGFBP) 2-6, but not IGFBP-1, are expressed in MDG2.1 cells. The secretion of IGFBPs from MDG2.1 cells shows > 10-fold levels of the 24 kDa form, reduced secretion of other IGFBPs, with no change in the total amount of IGFBP secreted, as compared to primary cells cultured under identical conditions. Use of endoglycosidase F indicated that several IGFBPs are posttranslationally modified. This cell line is a useful model and plasmid transfection target system to investigate IGFBP action in ovarian granulosa cells.

Animals↗
PubMed1993

Anaplastic large-cell Ki-1 lymphoma. Pulmonary presentation mimicking miliary tuberculosis.

An unusual presentation of anaplastic large-cell (ALC) Ki-1 lymphoma is described, in which pulmonary involvement mimicked miliary tuberculosis. Lung involvement is uncommon in this type of lymphoma, and it is not clear from previous reports whether this was a presenting feature. Bone marrow involvement, rare in Ki-1 lymphoma, also was present in this case. Despite therapy skin involvement subsequently developed, and the patient died 3 months after presentation. Postmortem examination revealed lymphomatous involvement of the heart, liver, kidneys, stomach, and lymph nodes. This case highlights the need for awareness of ALC Ki-1 lymphoma particularly when it presents at uncommon extranodal sites.

Adult↗
PubMed1993

Divergent actions of prostaglandins-E2 and -F2 alpha on the regulation of insulin-like growth factor-binding protein-3 in luteinized granulosa cells.

Insulin-like growth factor (IGF)-I synergizes with gonadotropin to further stimulate ovarian steroidogenesis. In contrast, the IGF-binding proteins (IGFBPs) produced by granulosa cells have been shown to antagonize the stimulatory actions of the IGFs and gonadotropins. The purpose of the present study was to examine the effects on IGFBP-3 production of prostaglandin (PG)-E2, a compound known to stimulate luteal function and prevent/delay luteal regression (a luteotropic compound), and PGF2 alpha, a compound known to be luteolytic. PGF2 alpha significantly stimulated IGFBP-3 production to 2.6-fold of control (P < 0.05) while PGE2 attenuated its production to half of control (P < 0.05). In contrast to the effects of IGFBP-3, PGE2 stimulated progesterone production to 8-fold of control (P < 0.05) while PGF2 alpha had no effect. Possible mechanisms of action of PGE2 and PGF2 alpha were also examined. PGE2, but not PGF2 alpha, stimulated cAMP accumulation which has been previously shown to inhibit IGFBP-3 production. PGF2 alpha is suspected to act via activation of protein kinase-C. However, a phorbol ester did not mimic PGF2 alpha's action toward IGFBP-3. This study demonstrated that PGE2 and PGF2 alpha conversely modulate IGFBP-3 production. Since IGFBPs have been shown to antagonize gonadotropin and IGF actions, this action of the prostaglandins may impact on the synergism between IGFs and gonadotropin necessary for follicular growth and luteal function.

1-Methyl-3-isobutylxanthine↗
PubMed1993

Expression of the mRNAs for the insulin-like growth factors and their binding proteins during development of porcine ovarian follicles.

We examined the expression of the mRNAs for the insulin-like growth factors (IGFs) and two of their binding proteins (BPs), IGFBP-2 and IGFBP-3, in individual follicles of the porcine ovary. Follicular development was synchronized with a progestin (altrenogest). Individual follicles were isolated on days 1, 3, 5 and 7 after progestin withdrawal. No IGFBP-3 mRNA was detected. While IGF-II mRNA was easily detected, the levels of expression did not change. IGF-I and IGFBP-2 mRNAs increased and decreased, respectively, with follicle development until day 7 when IGF-I expression declined. Regression analysis of IGF-I and IGFBP-2 mRNA expression was performed to assess the relative strength of correlations with day, diameter and steroid concentrations as covariates. IGFBP-2 mRNA was correlated with both day and diameter (r = -.713 and -.705, respectively, n = 24) and neither estrogen (E2) nor progesterone (P4) contributed to the fit. While IGF-I mRNA expression was correlated to both day (r = .483) and diameter (r = .587), the strongest predictor was E2 concentration (r = .694, n = 27). In conclusion, the expression of IGF-I and IGFBP-2 mRNAs in the ovarian follicle are discordantly regulated during follicular growth and maturation. The observed changes in these parameters should result in increased bioavailable IGF-I. This supports a pivotal autocrine/paracrine role for these factors during follicle growth and development.

Animals↗