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Biomedical subjects

J M Hammond

Publications and source records attributed to J M Hammond.

At least 37 records · Page 2Linked to original sources
PubMed1997

Intensive care management of varicella pneumonia.

To determine the clinical features, treatment and outcome of severe varicella pneumonia with hypoxic respiratory failure requiring intensive care management, a prospective survey of consecutive cases was undertaken. Fifteen consecutive adult cases of varicella pneumonia with respiratory failure admitted to a 10-bed respiratory intensive care unit over a period of 10 y from 1984-1993 were studied. All patients were given acyclovir immediately on admission. The level of ventilatory support needed was determined by the severity of gas exchange abnormality, and varied from face mask oxygen (three patients), through continuous positive airway pressure (CPAP) by face mask (eight patients), to continuous positive pressure ventilation (CPPV) (four patients). The majority of patients were young females, only one of whom was pregnant. All patients had been in close contact with a known case of chickenpox. All patients responded well to acyclovir and ventilatory support with improved oxygenation. Monitoring with pulse oximetry was important to detect episodes of desaturation on inadvertent discontinuation of positive and expiratory pressure (PEEP). Two patients were admitted with bacterial superinfection, and one patient, who had required intubation and CPPV, developed nosocomial respiratory tract infection. There were no deaths. This experience suggests that intensive care admission, with the early administration of intravenous acyclovir and recognition of the severity of the hypoxaemia resulting from varicella pneumonia (which can be reversed with PEEP), should reduce the mortality of severe varicella pneumonia in adults.

Acyclovir↗
PubMed1996

The intensive care management, mortality and prognostic indicators in severe community-acquired pneumococcal pneumonia.

OBJECTIVE: To determine mortality and factors that might predict outcome in severe community-acquired pneumococcal pneumonia treated by a standard protocol. DESIGN: Prospective, non-concurrent study. SETTING: Respiratory intensive care unit (ICU) in a teaching hospital. PATIENTS: 63 patients who were diagnosed by positive blood culture or Gram stain and culture of sputum or tracheal aspirate were included. MEASUREMENTS AND RESULTS: Clinical features, severity scores including Acute Physiology and Chronic Health Evaluation (APACHE) II, organ failure and lung injury scores, and the clinical course in the ICU were documented; 79% of patients required mechanical ventilation. Bacteraemia was present in 34 patients (54%); there were no distinguishing clinical features between bacteraemic and non-bacteraemic cases. The overall mortality was 21%, with only 5 deaths (15% mortality) in the bacteraemic group. Shock and a very low serum albumin (< 26 g/l) were the only clinical features that differentiated survivors from non-survivors; lung injury, APACHE II and multiple organ failure scores were all predictive of outcome. The positive predictive value and specificity in predicting death in individuals for the modified British Thoracic Society rule 1 were 26 and 64%; APACHE II > 2057 and 88%; > 2 organ failure 64 and 92%; and lung injury > 233 and 73%, respectively. CONCLUSIONS: These results suggest that even in bacteraemic cases mortality should be below 25% with intensive care management and that conventional scoring systems, while predictive of group mortality, are unreliable in individuals.

Adult↗
PubMed1996

Postsynthetic regulation of insulin-like growth factor-binding protein-3 by MCF-7 human breast cancer cells in culture.

Breast cancer cells are exposed to insulin-like growth factors (IGFs) which stimulate their proliferation, and to IFG-binding proteins (IGFBPs) which sequester and modulate IGF action. The primary circulatory IGFBP is IGFBP-3. In the present study, cultured MCF-7 breast cancer regulated clearance of IGFBP-3 via both cell association and proteolysis. Exogenously added IGFBP-3 was significantly cleared from the medium over time yielding the formation of smaller sized immunodetected fragments. Clearance was inhibited by IGF-I and -II. In contrast, clearance was not affected by growth factors and an IGF-analog having mitogenic activity but not binding to IGFBPs. In fact, activity of the IGFs and analogs paralleled their degree of binding to the IGFBP, suggesting that the IGF-binding altered IGFBP-3 making it less susceptible to clearance. Qualitatively similar results were obtained when these experiments were conducted using cell-free conditioned medium, thus suggesting the presence of secreted protease(s). However, level of proteolytic activity was much less than that found in the presence of cells. Clearance of rhIGFBP-3 also involved binding to the cell. Disappearance of rhIGFBP-3 was shown to be attenuated by heparin, which blocks cell surface binding sites. In contrast, compounds which block internalization did not inhibit IGFBP-3 clearance.

Breast Neoplasms↗
PubMed1996

Expression of the messenger ribonucleic acids for insulin-like growth factor-I and insulin-like growth factor binding proteins in porcine corpora lutea.

In the pig, the corpus luteum (CL) can develop and function autonomous of pituitary gonadotropins for approximately 12 days. We hypothesized that the insulin-like growth factor (IGF) system may play an autocrine/paracrine luteotrophic role(s) during this period. In this study, we monitored the expression (i.e., steady-state levels of mRNAs) of IGF-I and IGF binding proteins (IGFBP)-2, -3, -4, -5, and -6 mRNAs in whole CL and in small and large luteal cells on Days 4-16 of the estrous cycle. CL were dissociated with collagenase, and large and small luteal cells were isolated by centrifugal elutriation. Whole CL and luteal cells were extracted to isolate total or poly(A)+ RNA, which was subjected to Northern and/or dot-blot analyses using [32P]-labeled cDNA probes for IGF-I and IGFBP-2, -3, -4, -5, and -6. Northern blots showed readily detectable transcripts for IGF-I (6.7 and 0.9 kb), IGFBP-2 (1.8 kb), IGFBP-3 (2.8 kb), IGFBP-4 (2.6 kb), and IGFBP-5 (6.0 kb), but not for IGFBP-6. IGFBP-3 and -5 transcripts were observed mainly in small luteal cells, while IGFBP-2 and -4 were seen in both cell types. Dot-blot analyses for IGF-I and IGFBP-3 mRNAs were performed on total RNA from small and large luteal cells; blots were counter-probed with 3-phosphoglyceraldehyde dehydrogenase (p-GAD) cDNA to assess RNA quantity and quality. IGF-I mRNA (ratio IGF-I:p-GAD mRNA) expression was approximately 2-fold greater in small than in large luteal cells on Days 4-10. However, steady-state levels of IGF-I mRNA in small, but not large, luteal cells decreased significantly on Days 12-16 (vs. Days 4-10). IGFBP-3 mRNA expression was significantly greater (approximately 3-fold) in small than in large luteal cells but did not vary significantly between Days 4-10 and 12-16 for either cell type. We conclude that porcine CL express mRNAs for IGF-I and IGFBP-2, -3, -4, and -5, and that while small luteal cells are the major sources of IGF-I and IGFBP-3 and -5, IGFBP-2 and -4 appear to be expressed to approximately the same extent in small and large luteal cells. These results further suggest that the IGF-I/IGF system may have autocrine/paracrine regulatory actions in CL development/function in the pig.

Animals↗
PubMed1996

The influence of surgery on cytokines in patients with intra-abdominal sepsis.

The cytokine cascade which is triggered by severe sepsis may contribute to progressive organ dysfunction and death from sepsis. This cascade may be accentuated by surgery for sepsis and pre-treatment with cytokine blockers could possibly ameliorate the response. This prospective controlled study determined the effect of surgery in 11 haemodynamically stable patients undergoing laparotomy for intra-abdominal sepsis. Serum levels of endotoxin, IL-1, IL-6, IL-8 and TNF-alpha were determined; blood cultures, features of systemic inflammatory response, and organ dysfunction were monitored over the peri-operative period. There was considerable variation in the serum cytokine levels. The preoperative IL-6 levels were significantly elevated in the septic patients and a threefold increase in IL-6 levels occurred in both groups postoperatively. An increase in TNF-alpha did not achieve significance because of high levels in control patients with cancer. Cytokine release which occurs during abdominal surgery is increased in patients with intra-abdominal sepsis.

Abdominal Abscess↗
PubMed1996

Regulation of insulin-like growth factor I biosynthesis in porcine granulosa cells.

In the present studies we examined the regulation of insulin-like growth factor I (IGF-I) expression in porcine granulosa cells in vitro. Using Northern analysis and ribonuclease protection assays with exon-specific probes, we identified the IGF-I messenger RNA (mRNA) transcripts present in these cells under basal and hormone-stimulated conditions. We also assessed changes in secreted IGF-I using Western blots and correlated the change in protein secretion after hormone treatment with changes in mRNA levels. By analogy to the human IGF-I gene and its transcription, two major transcripts of approximately 1 and 7.5 kilobases, seen in freshly isolated granulosa cells and follicle wall and in single passaged granulosa (MDGp1) cells, most likely correspond to IGF-IA. Minor transcripts of 3-4 kilobases, which appeared after FSH or forskolin treatments or in control cells after long exposure of the autoradiographs, were attributed to incompletely processed RNA precursors. Ribonuclease protection assay analysis using probes to detect alternative use of exon 5 or exon 6 indicated that most, if not all, of the transcripts contained only exon 6 sequence (IGF-IA). Both class 1 and class 2 transcripts were identified using exon 1- and exon 2-specific probes, respectively. GH increased steady state levels of IGF-I mRNA 3-fold, FSH increased it approximately 10-fold, and forskolin maximally increased it 12- to 15-fold. Estradiol had no effect alone or in combination with the other treatments. All treatments that increased IGF-I mRNA coordinately increased both class 1 and class 2 transcripts, with the increase in class 1 greater than that in class 2. Multiple forms of IGF-I protein were seen under basal conditions and after hormone treatment. These were identified based on mRNA analysis and biochemical methods as both glycosylated and nonglycosylated IGF-IA prohormone, incompletely processed forms of prohormone, and the mature peptide. Changes in the levels of total protein were similar to the changes in mRNA (GH, 3-fold; FSH and forskolin, 10- to 20-fold). All forms of the protein changed coordinately, suggesting that these hormones had no major effect on the intracellular processing mechanism. IGF-binding protein-3 was able to bind to all IGF-I forms. These data conclusively demonstrate FSH and GH induction of ovarian IGF-I. The porcine granulosa cell culture system used in these studies should be an excellent system for studying the hormonal regulation of IGF-I expression.

Animals↗
PubMed1996

Selective expression of insulin-like growth factor system components during porcine ovary follicular selection.

In situ hybridization was used to map anatomical patterns of insulin-like growth factor (IGF) system gene expression in the gilt ovary through the estrous cycle. IGF-I, IGF-II< and IGF-I receptor messenger RNAs (mRNAs) were coexpressed in granulosa cells of developing and dominant follicles through the follicular phase. IGF-I and IGF-I receptor mRNAs were selectively concentrated in healthy follicles, whereas IGF-II mRNA was found in all follicles regardless of incipient or overt atresia. IGF-binding protein-1 (IGFBP-1) was not detected in the gilt ovary at any stage. IGFBP-2 mRNA was most abundant in granulosa cells of small follicles and in the ovarian vasculature regardless of cycle stage. IGFBP-2 mRNA persisted in granulosa cells of preovulatory follicles and corpora lutea. IGFBP-3 mRNA was not detected in developing follicles, but was detected in all luteal phase corpora lutea, apparently expressed by theca lutein cells. IGFBP-4 demonstrated a highly dynamic pattern of gene expression, closely tracking LH receptor gene expression throughout the follicular and luteal phases of the estrous cycle. IGFBP-4 mRNA was concentrated in the theca of medium to large growing follicles, but was not detected in granulosa cells until the emergence of dominant follicles in the late follicular stage, when the granulosa cells showed morphological evidence of luteinization as well as LH receptor gene expression. IGFBP-4 mRNA expression continued in granulosa cells of corpora lutea during the luteal phase and was not detected in atretic follicles. IGFBP-5 mRNA was concentrated in the surface or germinal epithelium and in capillary endothelium, particularly in capillaries of corpora lutea. In summary, there is selective expression of IGF-I, IGF-I receptor, and IGFBP-2 and -4 mRNAs during the process of follicular selection in the gilt ovary, with IGFBP-4 expression being closely associated with follicular selection and luteinization. These observations support an important role for autocrine/paracrine IGF action modulated by IGFBP-2 and -4 in both follicular growth and differentiation in the porcine ovary.

Animals↗
PubMed1995

Expression of cytochrome P450 1A1, an estrogen hydroxylase, in ovarian granulosa cells is developmentally regulated.

In this paper we report the analysis of porcine ovarian granulosa cells for the expression of several known hepatic estrogen hydroxylase RNAs. Of the P450s examined, only CYP 1A1 RNA was detected. Accordingly, the regulation of this mRNA was studied. The RNA for CYP 1A1 was dramatically and completely induced within 2 hours after exposure of immortalized granulosa cells to 3-methyl-cholanthrene (3MC) and expression could be inhibited with 10 microM phorbol myristate acetate. This message was also inducible by 3MC in cultured primary granulosa cells isolated from immature and developing follicles. Dexamethasone increased the relative expression of CYP 1A1 RNA in 3MC treated cells. In the absence of 3MC, the CYP 1A1 message was expressed in cultured granulosa cells from developing but not immature follicles, indicating developmental regulation of this enzyme. Further support for developmental regulation was provided by studies which detected the appearance of CYP 1A1 RNA during growth of ovarian follicles in vivo. This is the first report identifying a specific P450 estrogen hydroxylase RNA in ovarian granulosa cells.

Animals↗
PubMed1995

Long-term effects of selective decontamination on antimicrobial resistance.

OBJECTIVE: To determine whether selective decontamination of the digestive tract exerts any long-term effects on antimicrobial resistance patterns. DESIGN: A surveillance and interventional study comparing the antimicrobial sensitivity patterns of clinically important bacterial isolates the year before a 2-yr, double-blind, randomized, controlled study of selective decontamination of the digestive tract, and for the year thereafter when no use of the regimen was made. SETTING: A ten-bed respiratory intensive care unit (ICU) in a 1,200-bed teaching hospital. PATIENTS: All 1,528 patients admitted to the ICU over the 4-yr study period were included. There were 406 patients admitted in the year before the study of decontamination of the digestive tract (65% medical, 23% surgical, and 12% trauma), of whom 76% required mechanical ventilation. There were 719 patients admitted during the 2-yr study of selective decontamination (55% medical, 28% surgical, and 17% trauma), of whom 79.6% required mechanical ventilation. There were 403 patients admitted in the subsequent year (61% medical, 25% surgical, and 14% trauma), of whom 76.9% required mechanical ventilation. INTERVENTIONS: We performed daily clinical monitoring to detect nosocomial infection, with microbiological investigation when clinically indicated, as well as twice-weekly routine microbiological surveillance sampling. Antimicrobial susceptibility testing using standard laboratory methods was also performed. Selective decontamination of the digestive tract included parenteral cefotaxime and oral and enteral polymyxin E, amphotericin B, and tobramycin. MEASUREMENTS AND MAIN RESULTS: The occurrence rate of nosocomial infection was 20.6%, 16.6%, and 25.3%, respectively, in the three study periods. In the year after selective decontamination, there was an increase in the occurrence rate of infection (p = .005), with an-associated increase in infections caused by the Enterobacteriaceae, while a reduction in the level of resistance to the third-generation cephalosporins were found (p = .07). There was a progressive increase in the occurrence rate of infections caused by Acinetobacter species (p = .05). Only 11 infections over the 4 yrs were caused by Enterococcus species. Staphylococcal infections were uncommon (5.7% of admissions), and the level of methicillin resistance did not change. No increase in aminoglycoside resistance occurred. CONCLUSION: No long-term effects on antimicrobial resistance or the spectrum of nosocomial pathogens could be attributed to the use of selective decontamination of the digestive tract over a 2-yr period in a respiratory ICU admitting all categories of patients.

APACHE↗
PubMed1995

Insulin-like growth factor binding protein-3 inhibits porcine granulosa cell function in vitro.

Recombinant human insulin-like growth factor binding protein-3 (rhIGFBP-3) effects on basal, insulin-like growth factor I (IGF-I)-, and follicle-stimulating hormone (FSH)-stimulated progesterone (P4) secretion and [3H]aminoisobutyric acid (AIB) uptake by primary porcine granulosa cells (MDGs) and MDGs that have been passaged once (MDGp1) were assessed. Cells were treated concurrently or were preincubated with rhIGFBP-3 followed by treatment. rhIGFBP-3 had no effect on MDG or MDGp1 cell numbers after 24 h. Cotreatment with rhIGFBP-3 inhibited P4 secretion after treatment with FSH, IGF-I, and FSH plus IGF-I. FSH did not stimulate [3H]AIB uptake. However, the IGF-I-stimulated increase in [3H]AIB uptake was completely prevented by concurrent treatment with IGFBP-3. Preincubation of MDGp1 cells with IGFBP-3 dose dependently inhibited FSH- and IGF-I-stimulated P4 secretion. This inhibition was associated with increased cell association of the binding protein and increased IGF-I binding to the cells. These results indicate that IGFBP-3 is inhibitory to a variety of crucial functions in porcine granulosa cells, supporting a role for it in the regulation of granulosa cell function.

Aminoisobutyric Acids↗
PubMed1995

Is there a role for selective decontamination of the digestive tract in primarily infected patients in the ICU?

The role of selective decontamination of the digestive tract (SDD) for the prevention of nosocomial infection in critically ill patients remains controversial, and the efficacy of this technique in patients who are already infected on presentation to the intensive care unit has not previously been assessed. We performed a double-blind randomized placebo controlled trial of SDD (parenteral cefotaxime, six-hourly oral and enteral polymyxin E, tobramycin, and amphotericin B vs placebo) for all infected patients presenting to the ICU requiring mechanical ventilation for more than 48 hours and ICU stay of more than 5 days. Daily clinical and microbiological monitoring for secondary infection was undertaken until hospital discharge. In all, 59 selective decontamination and 76 placebo fully comparable patients fulfilled criteria for enrollment and analysis (APACHE II 15.2 vs 15.1). The number of patients receiving SDD who developed nosocomial infections was significantly reduced (P = 0.048), and there were no infections caused by the enterobacteriaceae or Candida spp in this group. No difference in ICU (17.5 vs 18.8 days) or hospital stay (32.7 vs 34.2 days) or mortality (17% vs 22.3%) was shown. Critically ill, primarily infected patients are protected from nosocomial infection by the use of SDD.

Administration, Oral↗
PubMed1995

The etiology and antimicrobial susceptibility patterns of microorganisms in acute community-acquired lung abscess.

OBJECTIVE: To determine the spectrum and antibiotic susceptibility patterns of microorganisms causing acute community-acquired lung abscess. DESIGN: A prospective survey. SETTING: Medical emergency department and wards of a tertiary teaching hospital. PATIENTS: Thirty-four adult patients with both clinical and radiologic features compatible with a diagnosis of acute community-acquired lung abscess who had received less than 48 h of antibiotic therapy. INTERVENTIONS: Microbiologic specimens obtained by percutaneous lung aspiration and with a protected specimen brush via fiberoptic bronchoscopy were submitted for aerobic and anaerobic culture. MAIN OUTCOME MEASURES: Identification of all microorganisms, including anaerobes, and determination of antibiotic susceptibility. RESULTS: A mean of 2.3 bacterial species per patient was isolated, anaerobes alone being isolated in 44% of cases, aerobes alone in 19%, and mixed aerobic and anaerobic isolates in 22%. Aerobic Gram-negative pathogens were uncommon. In seven patients, Mycobacterium tuberculosis was identified; in two it was associated with other bacteria. In four patients, no organisms were isolated. All the nonmycobacterial isolates were susceptible to amoxicillin-clavulanate and in addition the anaerobes were all susceptible to chloramphenicol and almost all to a combination of penicillin and metronidazole. Among the anaerobes, the level of resistance to penicillin, metronidazole, and clindamycin individually was 21%, 12%, and 5%, respectively. CONCLUSIONS: Community-acquired acute lung abscess is usually caused by multiple anaerobic and less frequently aerobic Gram-positive microorganisms, which should respond to empirical therapy with amoxicillin-clavulanate, chloramphenicol, or a combination of penicillin and metronidazole. Tuberculosis, which may be indistinguishable from an acute lung abscess, occurred in 21% of patients in our study. Most bacterial pathogens are sensitive to conventional antimicrobial therapy and further investigation with percutaneous lung aspiration or bronchoscopy is indicated only when there is lack of early response to therapy or there is the presence of atypical clinical features.

Acute Disease↗
PubMed1995

Changes in insulin-like growth factor-binding protein-2 and -3 in follicular fluid during atresia of follicles grown after ovulation in pigs.

In pigs, medium-sized follicles (3-6 mm in diameter), lost during preovulatory maturation, are rapidly replenished after ovulation, increasing tenfold in number between day 3 and day 8 of the oestrous cycle. The incidence of atresia in medium-sized follicles was previously found to increase between day 5 and day 7 after the onset of oestrus (day 0). This study was conducted to determine whether changes in concentrations of insulin-like growth factor binding proteins (IGFBPs) in pig follicular fluid from medium-sized follicles were related to increased incidence of atresia. Individual follicles were isolated on days 5, 6 and 7 after oestrus from eleven pigs. Follicles were defined as atretic when they contained > or = 10% apoptotic granulosa cells (cells containing subdiploid amounts of DNA fluorescence determined by flow cytometry). Concentrations of IGFBP-2 and -3 were determined by ligand blot analysis with 125I-labelled insulin-like growth factor II and quantified by densitometry. The mean percentage of apoptotic cells per follicle increased (P < or = 0.01) from 3.6 to 35.0 and the mean concentration of IGFBP-2 increased (P < or = 0.001) 3.3-fold between day 5 and day 7 after oestrus. The concentration of IGFBP-3 did not differ significantly among days. Mean oestradiol concentration in follicular fluid decreased (P < or = 0.05) from 52.3 to 0.9 ng ml-1 between day 5 and day 7. Androstenedione concentrations in follicular fluid also decreased (P < or = 0.01) between day 5 and day 7, but progesterone did not differ significantly among days.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstenedione↗
PubMed1995

Prophylactic use of the new quinolones for prevention of nosocomial infection in the intensive care unit.

The new quinolone antimicrobial agents, particularly those with less activity against anaerobes, selectively prevent colonisation of the alimentary tract by Gram-negative bacilli and staphylococci without substantially affecting the normal anaerobic flora, which preserve the colonisation resistance of the gut. These properties ideally position this class of antibacterial agent for selective decontamination of the digestive tract (SDD) in the prevention of nosocomial infection. The rationale for this procedure is based on the presumption that a significant proportion of infections in compromised patients are endogenous in origin, arising from the host's own microbial flora. If this colonisation by potentially pathogenic microflora within the normal flora can be significantly reduced without being replaced by other more pathogenic microorganisms, the risk of endogenous infection should be minimised. The quinolones have proved to be ideal agents for use in preventing infection in bone marrow transplant and other neutropenic patients. They have been used for SDD in the general intensive care unit population, although the technique has not received widespread acceptance. There have been only 4 reported randomised studies using quinolones as part of SDD regimens and only 301 patients have been evaluated. Although the incidence of ventilator-associated pneumonia has been significantly reduced from 36 to 15%, no effect has been shown on mortality. The cost of using SDD is significantly less with the quinolones than with other regimens, and induction of resistance has not been noted. The new quinolones, and in particular the more recently developed agents with extended Gram-positive activity, appear to be ideally suited for SDD, and their careful evaluation in further large, well designed trials is warranted.

4-Quinolones↗
PubMed1995

Plasma growth hormone, insulin-like growth factor I, and insulin-like growth factor binding proteins in pigs with divergent genetic merit for postweaning average daily gain.

Characteristics of growth hormone (GH), IGF-I, and IGF-binding proteins (IGFBP) were studied in gilts sampled from lines of pigs selected for either fast (line F, n = 14) or slow (line S, n = 14) postweaning ADG. Repeated blood samples were obtained from gilts (approximately 55 kg BW) during a period of feed deprivation and again during refeeding. Averaged across time, the difference in mean plasma GH concentrations of F and S gilts was not significant (7.7 vs 6.4 ng/mL; P > .20) during feed deprivation, and frequently, height, and amplitude of GH pulses did not differ (P > .25) for F and S pigs. Overall, F gilts had greater concentrations of plasma IGF-I than S gilts during feed deprivation (217.3 vs 145.1 ng/mL; P < .03). Across line, plasma IGF-I decreased (P < .01) during feed deprivation. Average GH did not differ (P > .40) for F and S gilts during the refeeding period. Average plasma IGF-I tended (P = .05) to be greater in F gilts than in S gilts during refeeding. Consistent with changes over time during feed deprivation, plasma IGF-I averaged across line increased (P < .01) in response to refeeding. Averaged across time (0 and 48 h refeeding), activity of IGFBP-2 (singlet band at 34 kDa) did not differ significantly (P = .17) in F and S gilts. However, there was a tendency (P = .13) for a line x time interaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Follicular atresia in pigs: measurement and physiology.

The physiological regulation of follicular atresia was investigated during the early luteal phase after ovulation and during altrenogest-synchronized preovulatory maturation in pigs (gilts). Apoptosis in dispersed granulosa cells was determined by flow cytometry. Apoptotic (A0) cells contain low, subdiploid amounts of DNA fluorescence. Follicles were classified biochemically as atretic or nonatretic based on the percentage of A0 (% A0) cells, atretic with > or = 10%, and nonatretic with < 10% A0 granulosa cells. The % A0 granulosa cells/follicle ranged from .02 to 89. Follicles containing debris in their isolated granulosa cells were classified as morphologically atretic. The morphological and biochemical criteria of atresia were in agreement for 224 of 248 follicles. Internucleosomal DNA cleavage, the hallmark of apoptosis, was determined by autoradiographic analysis of [32P]3'-end labeled DNA from granulosa cells. Densitometric analysis showed that optical density of [32P]3'-end labeled DNA fragments in the .18 to 20 kbp size range was correlated with the % A0 cells (R > .9, n = 22, P < .001). During altrenogest-synchronized preovulatory maturation, < 5% of large (> 6 mm in diameter) follicles were atretic. Among medium-sized follicles (3 to 6 mm) on d 1 and 3 of preovulatory maturation, only 17% were atretic, in contrast with d 5 when 87% were atretic. During the early luteal phase, atretic follicles/pig increased from 6% on d 5 to 50% on d 7 after estrus. Follicular fluid estradiol-17 beta concentration was greater (P < .001) in nonatretic than in atretic follicles on d 5 and 6 after estrus, but by d 7 estradiol-17 beta had decreased to a mean < 1 ng/mL in nonatretic and atretic follicles. The increase in apoptosis in granulosa cells and loss of estradiol-17 beta production in vivo indicated a high incidence of atresia among the first group of follicles grown after ovulation in pigs. These results indicate that apoptotic cell death was involved in degeneration of granulosa cells and atresia during two different stages of follicular development.

Animals↗
PubMed1994

Use of a recombinant antigen in an indirect ELISA for detecting bovine antibody to capripoxvirus.

The gene coding for the capripoxvirus structural protein P32 was cloned, expressed in Escherichia coli as a fusion protein with glutathione-S-transferase, and purified on glutathione Sepharose. An indirect enzyme linked immunosorbent assay (ELISA) using this antigen was developed to screen bovine sera for antibodies to capripoxvirus. Sequential serum samples from experimentally infected animals tested by ELISA and by virus neutralisation test (VNT) showed that the ELISA was more sensitive and detected antibodies to capripoxvirus earlier post-infection than the VNT.

Animals↗