ABSTRACT
Respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infection in infants. Winter outbreaks in Chile result in 5% of infected children hospitalized, with 0.01% mortality. Increased evidence indicates that viral and host factors modulate the severity of infection. Using DNA microarrays, we characterized the genome-wide transcriptional response of lung mucoepidermoid cells (NCI-H292) at 0, 24, 48, 72 and 96 hours post-infection (hpi) with aâ¯single dose of RSV/A. During the whole studied period, aâ¯bi-phasic gene expression profile was observed by aâ¯total of 330 differentially expressed genes. About 60% of them were up-regulated between 24-72 hpi and then turned-off at 96 hpi. This transient, early gene expression pattern was significantly enriched in biological processes like interferon signaling, antigen processing and presentation, double-stranded RNA binding and chemokine activity. We detected 27 common genes up-regulated between 24-72 hpi, from which IFIT1, IFI44, MX1, CXCL11 and OAS1 had the highest expression. The second pattern comprised over 120 genes, which remained silenced until 72 hpi, but were steeply up-regulated by 96 hpi. Biological processes of this late-response profile included cell cycle division and microtubule cytoskeleton organization. Conversely, the genes belonging to virus response pathway showed aâ¯decreased expression at 96 hpi. We conclude that RSV induces an early innate immune activation profile response until 72 hpi. Thereafter, the viral response is inhibited, leading to host cell recovery. The presented cellular model allows to study the specific pathways involved in elimination of infection at prolonged time intervals and their subsequent analysis in severe RSV disease of infants and/or older adults.
Subject(s)
Epithelial Cells/metabolism , Respiratory Syncytial Virus Infections/genetics , Respiratory Syncytial Virus, Human/physiology , Epithelial Cells/virology , Gene Expression Profiling , Humans , Lung/metabolism , Lung/virology , Respiratory Syncytial Virus Infections/metabolism , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Human/genetics , Respiratory Syncytial Virus, Human/isolation & purification , Time Factors , TranscriptomeABSTRACT
Genetically reducing boar taint using low-taint lines is considered the most sustainable and economic long-term alternative to surgical castration of male pigs. Owing to the high heritability of the main boar taint components (androstenone, skatole and indole), breeding is an excellent tool for reducing the number of tainted carcasses. To incorporate boar taint into breeding programmes, standardized performance testing is required. The objective of this study was to develop and formally present a performance test for the main boar taint compounds on live breeding candidates. First, a standardized performance test for boar taint was established. A biopsy device was developed to extract small tissue samples (200 to 300 mg) from breeding candidates. Quantification of boar taint components from these small samples using specialized chemical extraction methods proved accurate and repeatable (r = 0.938). Following establishment of the method, biopsy samples of 516 live boars (100 to 130 kg live weight) were collected in the second step. Various mixed linear models were tested for each boar taint compound; models were ranked in terms of their information content. Pedigree information of 2245 ancestors of biopsied animals was included, and genetic parameters were estimated using univariate and multivariate models. Androstenone (in µg/g liquid fat (LF): mean = 0.578, σ = 0.527), skatole (in µg/g LF: mean = 0.033, σ = 0.002) and indole (in µg/g LF: mean = 0.032, σ = 0.002) levels obtained by biopsy were plausible. Heritability estimates for androstenone calculated with univariate (0.453) and multivariate (0.452) analyses were comparable to those in the literature. Heritabilities for skatole (0.495) and indole (0.550) were higher than that for androstenone. Genetic and phenotypic correlations were similar to those published previously. Our results show that data on boar taint compounds from small adipose samples obtained by biopsy provide similar genetic parameters as that described in the literature for larger samples and are therefore a reliable performance test for boar taint in live breeding candidates.
Subject(s)
Androsterone/metabolism , Indoles/metabolism , Skatole/metabolism , Swine/genetics , Swine/physiology , Adipose Tissue/metabolism , Animals , Breeding , Male , Pedigree , Specimen HandlingABSTRACT
To assess whether plasma prolactin (PRL) characteristics relate to lactogenesis and absence or presence of menstrual cycles, we measured bioactive PRL (BIO-PRL) using the Nb2 assay, immunoreactive PRL (IR-PRL) by radio-immunoassay, calculated equations describing the BIO-PRL-IR-PRL relationship and separated charged PRL isoforms (by chromatofocusing) in five amenorrhoeic and five cycling nursing women at 6 months postpartum and in 10 cycling non-nursing women. Plasma samples were drawn before and 30 min after a suckling episode at 0800, 1600 and 2400 h in nursing women and at the same hours in non-nursing women. BIO-PRL and IR-PRL concentrations were highest in amenorrhoeic nursing women, intermediate in cycling nursing women and lowest in cycling non-nursing women. The BIO-PRL-IR-PRL relationship shows that a given amount of IR-PRL corresponds to equivalent amounts of BIO-PRL in cycling nursing and cycling non-nursing women, and to a larger extent in amenorrhoeic nursing women. IR-PRL was present in plasma as several charge isoforms. Bioactive isoforms eluting at pH 6.0-5.1 were found in amenorrhoeic and cycling nursing women, reaching similar concentrations after suckling. Bioactive isoforms eluting at pH 7.0-6.1 were found only in amenorrhoeic nursing women. We speculate that isoforms eluting at pH 6.0-5.1 may play a role in lactation and isoforms eluting at pH 7.0-6.1, in lactational amenorrhoea.
Subject(s)
Lactation/physiology , Prolactin/physiology , Adult , Breast Feeding , Female , Humans , Menstrual Cycle/physiology , Protein Isoforms/physiologyABSTRACT
To assess whether the duration of lactational amenorrhoea can be predicted in individual women, we studied the pre- and post-suckling concentrations of immune prolactin (IR-PRL) and of bioactive prolactin (BIO-PRL) and basal concentrations of oestradiol in ten amenorrhoeic fully nursing women at 3 months post-partum. The women were of similar age, weight and had infants of similar growth rate. Five of these women were to experience long amenorrhoea (>180 days) and the others short amenorrhoea (<180 days). Blood samples were drawn 30 min after a suckling episode initiated at 0800 h, 1600 h and 2400 h. BIO-PRL distinguished between groups of women at 0030 h but not at other times, while there was considerable overlap between values for IR-PRL and oestradiol at all times studied. At 1630 h, the ratios post-suckling BIO-PRL: oestradiol and post-suckling IR-PRL:oestradiol were above 2000 in the women that were to experience long amenorrhoea and below this threshold in the other women. The ratio post-suckling BIO-PRL:oestradiol provided more information since the difference between the lowest ratio in the long amenorrhoea and the highest ratio in the short was 699, while it was 520 for the IR-PRL:oestradiol ratio. The determination of these ratios may help to predict the duration of lactational amenorrhoea in individual fully nursing women.
Subject(s)
Amenorrhea/blood , Estradiol/blood , Postpartum Period , Prolactin/blood , Adult , Breast Feeding , Female , Humans , Time FactorsABSTRACT
The inhibitory actions of prolactin on gonadal steroidogenesis have been reported in different species and under a variety of experimental approaches. In this study, the mechanisms of the in-vitro effects of human prolactin (hPRL) on human follicle stimulating hormone (hFSH)-induced aromatase activity were determined using cultured granulosa cells from diethylstilboestrol (DES)-primed immature rats. Human PRL caused a dose-dependent decrease in hFSH-induced 17 beta-oestradiol production, even when cells were cultured in the presence of a cAMP analogue (8-Br-cAMP). These effects of hPRL appeared to be specific, since addition of an anti-rat PRL receptor monoclonal antibody (mAb) mimicked the hPRL inhibitory effect upon steroidogenesis in rat granulosa cells. In order to assess the importance of tyrosine kinase and protein kinase-C activation in the hPRL inhibitory effects upon oestrogen biosynthesis, cells were cultured in the presence of kinase inhibitors. The results showed that addition of genistein or staurosporine (a tyrosine kinase and protein kinase-C antagonist respectively) to cultured granulosa cells resulted in potent inhibition of hPRL actions upon hFSH-induced aromatization in a dose-dependent manner. These observations suggest that tyrosine kinase and protein kinase-C activation are involved in the biochemical events leading to hPRL inhibitory effects at the gonadal level.
Subject(s)
Aromatase/biosynthesis , Follicle Stimulating Hormone/antagonists & inhibitors , Granulosa Cells/drug effects , Prolactin/pharmacology , Protein Kinase C/physiology , Protein-Tyrosine Kinases/physiology , 8-Bromo Cyclic Adenosine Monophosphate/pharmacology , Animals , Aromatase/genetics , Cells, Cultured , Culture Media, Conditioned/chemistry , Cyclic AMP/analysis , Enzyme Induction/drug effects , Enzyme Inhibitors/pharmacology , Estradiol/analysis , Female , Genistein , Granulosa Cells/enzymology , Isoflavones/pharmacology , Phosphorylation/drug effects , Protein Kinase C/antagonists & inhibitors , Protein Processing, Post-Translational/drug effects , Protein-Tyrosine Kinases/antagonists & inhibitors , Rats , Rats, Wistar , Staurosporine/pharmacologyABSTRACT
In our population, only half of fully nursing women remain amenorrheic 6 months postpartum. The other half recover their menstrual cycles between 90-180 days postpartum in spite of a high suckling frequency and elevated immunoreactive PRL (IR-PRL) concentrations. To further investigate the association of PRL with the recovery of ovarian function, we compared PRL bioactivity (BIO-PRL) 3-4 months postpartum in fully nursing amenorrheic women who subsequently experienced long (> 180 days; n = 5) or short (< 180 days; n = 5) lactational amenorrhea. In the present study, BIO-PRL in plasma was measured by the Nb2 lymphoma cell assay in samples taken before and 30 min after a suckling episode at 0800, 1600 and 2400 h. Women in the long amenorrhea group had higher overall mean BIO-PRL (mean +/- SE, 129.9 +/- 12.1 micrograms/L) than nursing women in the short amenorrhea group (66.6 +/- 5.2 micrograms/L; P < 0.05). Mean basal values were similar, but the women in the long amenorrhea group had more BIO-PRL in response to suckling (160.1 +/- 4.0 vs. 71.9 +/- 6.7 micrograms/L; P < 0.05). Compared with their respective basal values, nursing women in the long amenorrhea group demonstrated increased BIO-PRL in response to suckling, whereas the other group did not. The relationships between BIO-PRL and IR-PRL were similar in the two groups of nursing women before suckling. However, after suckling, the long amenorrhea group had significantly higher BIO-PRL levels than IR-PRL levels (P < 0.05, by likelihood test) than the short amenorrhea group. This suggests that suckling differentially changes in each group either the composition of PRL present or substances that may modify the bioactivity of PRL in plasma.
Subject(s)
Amenorrhea/blood , Amenorrhea/etiology , Lactation , Prolactin/blood , Adult , Biological Assay , Female , Humans , Immunologic Techniques , Time FactorsABSTRACT
Fifty nine sporadic cases and forty five cases from six outbreaks of salmonellosis occurring in Mendoza, Argentina between 1972-76 are reported. All 104 patients were studied epidemiologically searching for the etiologic agent, implicated food and contacts. Stools of patients and contacts were examined. Other clinical specimens and the implicated foods were examined bacteriologically. The Salmonella isolates were classified in eleven serotypes with the following order of frequency: a) Outbreaks: S. typhimurium (50,0%), S. derby (16,7%), S. newport (16,7%), S. bredeney (16,7%), S. enteritidis (16,7%), S. cholerae-suis (16,7%) and S. oranienburg (16,7%). b) Sporadic cases; S. typhimurium (35,9%), S. newport (15,6%), S. anatum (7,8%), S. oranienburg (6,2%), S. derby (4,7%), S. java (3,1%), S. cholerae-suis (3,1%), S. bredeney (1,6%), S. enteritidis (1,6%), S. minnesota (1,6%), S. urbana (1,6%), and Salmonella spp (17,2%). These results are compared with those obtained in the same areas between 1962-71 and with the serotype frequencies from different sources of infection found in Mendoza and other regions.
Subject(s)
Disease Outbreaks/epidemiology , Salmonella Food Poisoning/epidemiology , Salmonella Infections/epidemiology , Salmonella/isolation & purification , Adolescent , Adult , Aged , Argentina , Child , Child, Preschool , Feces/microbiology , Female , Food Microbiology , Gastroenteritis/microbiology , Humans , Infant , Male , Middle Aged , Serologic TestsABSTRACT
Fifty nine sporadic cases and forty five cases from six outbreaks of salmonellosis occurring in Mendoza, Argentina between 1972-76 are reported. All 104 patients were studied epidemiologically searching for the etiologic agent, implicated food and contacts. Stools of patients and contacts were examined. Other clinical specimens and the implicated foods were examined bacteriologically. The Salmonella isolates were classified in eleven serotypes with the following order of frequency: a) Outbreaks: S. typhimurium (50,0
), S. derby (16,7
), S. newport (16,7
), S. bredeney (16,7
), S. enteritidis (16,7
), S. cholerae-suis (16,7
) and S. oranienburg (16,7
). b) Sporadic cases; S. typhimurium (35,9
), S. newport (15,6
), S. anatum (7,8
), S. oranienburg (6,2
), S. derby (4,7
), S. java (3,1
), S. cholerae-suis (3,1
), S. bredeney (1,6
), S. enteritidis (1,6
), S. minnesota (1,6
), S. urbana (1,6
), and Salmonella spp (17,2
). These results are compared with those obtained in the same areas between 1962-71 and with the serotype frequencies from different sources of infection found in Mendoza and other regions.
