Biomedical subjects

J L Patterson

Publications and source records attributed to J L Patterson.

At least 19 recordsLinked to original sources

PubMed2005

Embryonic and fetal development in a commercial dam-line genotype.

During depopulation of a breeding unit within Swine Graphics Enterprises, extensive data were collected and used to examine relationships among ovulation rate, the pattern of prenatal loss, and placental and fetal development. Groups of Large White x Landrace females (n=447) were slaughtered between day 20-30, 50-55 or 85-90 of gestation, with approximately equal numbers of animals representing gilts and parity 1 (G/P1), parity 2-3 (P2/3), and parity >4 (P4+). Ovulation rate and embryo number were recorded for all animals. With the exception of the G/P1 animals, embryonic and placental weight were recorded for four conceptuses per sow on day 20-30; on day 85-90 two conceptuses per sow were dissected to determine placental and fetal development. Ovulation rate (22.7 +/- 0.2 overall) was higher (P <0.05) in P2/3 (23.6 +/- 0.4) and P4+ (24.7 +/- 0.4) than in G/P1 (20.2 +/- 0.5). Embryonic/fetal survival was 61.8 +/- 2.1% at day 20-30, 50.2 +/- 2.2% at day 50-55 and 48.7 +/- 1.9% at day 85-90 and the number of surviving conceptuses was higher (P <0.05) in the P2/3 sows than in other parity groups. There was no relationship between ovulation rate and number of live embryos at day 20-30 or 85-90. At day 20-30 and 85-90, embryo weight was positively correlated with placental weight, but neither placental weight nor embryonic/fetal weight was correlated with number of viable embryos. A parity by gestation day interaction existed; placental weight for P4+ (3.42 +/- 0.43 g) was less than for P2/3 (7.55 +/- 0.40 g) at day 20-30 (P <0.0001), whereas at day 85-90, placental weight of P2/3 (209.5 +/- 8.5 g) was less (P=0.05) than both G/P1 (235.7 +/- 7.3g) and P4+ (235.4 +/- 7.1 g). At day 85-90, fetal brain weight, relative to body weight (R2=0.61, P <0.0001), and fetal brain:liver weight ratio (R2=0.35; P <0.0001) were negatively related to mean fetal weight, and brain:liver weight ratio showed a trend towards a relationship with number of viable fetuses (P=0.08). Parity also affected brain:liver weight ratio (P=0.01). Clearly, high ovulation rates in the higher parity sows have the potential to cause excessive in utero crowding of conceptuses in the post-implantation period. Even with moderate crowding, increased brain:liver weight ratios in smaller fetuses in late gestation indicate that uterine capacity impacts fetal development as well as the number of surviving fetuses.

Animals↗

PubMed2002

Impact of boar exposure on puberty attainment and breeding outcomes in gilts.

We examined the most effective method of boar exposure for the attainment of puberty in 89 gilts. At 160 days of age, we allocated gilts to daily direct contact with a vasectomized boar after movement of pen groups of gilts to a detection-mating area (DGB: n = 30); daily direct contact with boars in the gilt home pens (DBG: n = 31); or daily fenceline contact between boars and gilts housed in individual gilt stalls (FBG: n = 28). DGB gilts were younger (P < or = 0.05) than FBG gilts at puberty. Direct boar contact reduced the interval from initial boar contact to puberty in DGB and DBG gilts, compared to fenceline contact in FBG gilts (P < 0.05). There was no difference (P > or = 0.05) between treatment for pubertal weight, backfat, lifetime growth rate, or duration of first pubertal estrus. Backfat depth and leptin concentration at 160 days of age were positively correlated (P < or = 0.05). We detected no relationships between leptin or IGF-1 concentration at 160 days of age and the interval from initial exposure to a vasectomized boar to puberty (P > 0.05). Based on objective criteria, fenceline contact with a boar (BC) during artificial insemination improved the quality of artificial insemination compared to no boar contact (NC) (P < 0.05).

Adipose Tissue↗

PubMed2002

The effect of lean growth rate on puberty attainment in gilts.

Two hundred sixteen prepubertal Genex Manor hybrid F1 gilts were used to determine the impact of lean growth rate on sexual development of gilts. This study was composed of two experiments (Exp. 1 and Exp. 2). In Exp. 1, at approximately 96 d of age and 54 kg weight, gilts were allocated with respect to growth rate and litter origin to one of two dietary treatments: 1) a diet formulated to maximize lean growth potential (LP; n = 84) or 2) a diet formulated to produce a lower lean growth rate (LL; n = 84). In Exp. 2, at approximately 88 d of age and 50 kg weight, gilts were allocated with respect to growth rate and litter origin to one of two dietary treatments: 1) a diet formulated to maximize lean growth potential (LP; n = 24) or 2) a diet formulated to restrict lean growth further than was achieved in LL in Exp. 1 (RL; n = 24). All gilts were fed treatment diets for ad libitum consumption and housed in groups of six. Weight, backfat depth and loin depth, and feed intake were measured weekly. Starting at 135 d of age, gilts received 20 min of direct daily exposure to a boar as a pen group for pubertal stimulation. Puberty attainment was determined as the day gilts first exhibited the standing reflex in response to contact with a boar. At pubertal estrus, body weight, backfat depth, and loin depths were recorded. Diet affected (P < or = 0.05) estimated fat-free lean gain (LP, 424 vs LL, 347 g/d, Exp. 1; LP, 397 vs RL, 376 g/d, Exp. 2) during the growth period (start to stimulation). However, age at puberty was not affected by diet (LP, 157.3 vs LL, 157.6, Exp. 1; LP, 166.7 vs RL, 167.3, Exp. 2) or overall lean growth at stimulation (P > or = 0.05 in both experiments), confirming that innate variability in sexual development of commercial genotypes, rather than growth performance, determines onset of sexual maturity. A negative correlation between age at puberty and growth rate from 50 kg until puberty (P < or = 0.05) (LP, r = -0.40, LL, r = -0.36, Exp. 1; LP, r = -0.64, RL, r = -0.48, Exp. 2) was a consequence of reduced lean tissue growth during the stimulation period in later-maturing gilts.

Adipose Tissue↗

PubMed2001

Lack of an effect of prostaglandin injection at estrus onset on the time of ovulation and on reproductive performance in weaned sows.

We evaluated the effects of a PGF2alpha analogue on time of ovulation and reproductive performance in multiparous Camborough sows (n=47). At onset of first post-weaning estrus, sows received either an intravulval injection of 3.75 mg of prostaglandin analogue (PGF) or, served as a non-injected control (CON). Beginning 24 h after the onset of estrus, transcutaneous ultrasonography was carried out every six h to determine time of ovulation. At 36, 54, and 72 h after the onset of estrus, blood samples were taken for progesterone analysis. Weaning-to-estrus (WEI), duration of estrus, ovulation rate and number of live embryos at d 28 of gestation were recorded. Treatment had no effect (P > 0.05) on any parameters measured. Duration of estrus classified as less or greater than the overall mean also had no effect (P > 0.05) on any of the parameters measured. Results indicate that treatment did not advance ovulation nor did it improve reproductive performance in sows. Overall, a negative correlation of WEI with the ovulation rate (P = 0.0005, r = -0.54) was established.

Animals↗

PubMed2000

Observed and reported expressive vocabulary and word combinations in bilingual toddlers.

The consistency of parental reports of expressive vocabulary and word combinations with observed expressive language among 21- to 27-month-old children exposed to English and Spanish on a regular basis was the focus of this investigation. Parental reports were obtained using the Spanish-English Vocabulary Checklist (Patterson, 1998), an adaptation of the Language Development Survey (Rescorla, 1989). The number of different words parents reported was correlated (r = .66) with the number of different words the children used during a 30-min videotaped interaction with the reporting parent. Parental reports of whether the child was combining words and estimates of proportion of the child's use of Spanish and English also were consistent with the children's language use during the 30-min language samples.

Child Language↗

PubMed2000

Identification of the minimal essential RNA sequences responsible for site-specific targeting of the Leishmania RNA virus 1-4 capsid endoribonuclease.

The Leishmania RNA virus 1-4 capsid protein possesses an endoribonuclease activity responsible for single-site-specific cleavage within the 450-nucleotide 5' untranslated region of its own viral RNA transcript. To characterize the minimal essential RNA determinants required for site-specific cleavage, mutated RNA transcripts were examined for susceptibility to cleavage by the virus capsid protein in an in vitro assay. Deletion analyses revealed that all determinants necessary for accurate cleavage are encoded in viral nucleotides 249 to 342. Nuclease mapping and site-specific mutagenesis of the minimal RNA sequence defined a stem-loop structure that is located 40 nucleotides upstream from the cleavage site (nucleotide 320) and that is essential for accurate RNA cleavage. Abrogation of cleavage by disruption of base pairing within the stem-loop was reversed through the introduction of complementary nucleotide substitutions that reestablished the structure. We also provide evidence that divalent cations, essential components of the cleavage reaction, stabilized the stem-loop structure in solution. That capsid-specific antiserum eliminated specific RNA cleavage provides further evidence that the virus capsid gene encodes the essential endoribonuclease activity.

Animals↗

PubMed1999

Host cell proteins bind specifically to the capsid-cleaved 5' end of Leishmaniavirus RNA.

Leishmaniavirus (LRV) is a double-stranded RNA (dsRNA) virus that persistently infects some strains of the protozoan parasite, Leishmania. LRV generates a short transcript, corresponding to the 5' end of the positive-sense RNA (320 nt), via a cleavage event mediated by the viral capsid protein on the full-length positive sense RNA transcript. To address the possibility that the RNA cleavage represents a regulatory mechanism for maintaining persistent infection, the interactions between Leishmania cytoplasmic proteins and in vitro synthesized viral transcripts were studied. In gel mobility shift experiments, three specific RNA/protein complexes were formed between cellular proteins and the cleaved viral transcript, and three major proteins were labeled by UV cross-linking. No protein binding activity was observed for either the short (320 nt) or full-length RNA transcripts. However, the two cleavage reaction products were able to form stable RNA/RNA complexes. We present a model in which the virus is targeting its own transcript for cleavage to promote binding of host factors to cryptic domains inaccessible in the full-length transcript.

5' Untranslated Regions↗

PubMed1998

Generation of the short RNA transcript in Leishmaniavirus correlates with the growth of its parasite host, Leishmania.

Leishmaniavirus 1 is a double-stranded RNA virus that infects the New World kinetoplastid parasites, Leishmania braziliensis, and Leishmania guyanensis. The isolated virus particles contain an RNA-dependent RNA polymerase which exhibits both transcriptase activity for genome-length plus-strand synthesis and replicase activity for genome-length minus-strand synthesis. Recently, we identified a 320 nucleotide short RNA transcript of Leishmaniavirus 1-4, derived from the 5' end of the viral plus-strand, which is generated by the virus capsid via site-specific cleavage of the full-length positive single-stranded RNA. We have hypothesized that this short RNA transcript functions to regulate the virus life cycle during the growth of its parasite host, Leishmania guyanensis. To address this hypothesis, we measured the relative amount of short RNA transcripts and the absolute number of viral genomes per infected cell from log through stationary phase of the parasite growth cycle. In vitro assays of the viral polymerase showed an overall increase in viral polymerase activity from log growth into stationary phase which mirrored an in vivo increase in the quantity of double-stranded genome as measured by agarose gel electrophoresis. We have developed competitive reverse transcription-polymerase chain reaction (RT-PCR) assays to measure the relative amounts of viral transcripts in infected cells as well as the number of viral genomes per infected cell. The results of these assays show that the amount of full-length virus transcripts peaks in the parasite stationary phase (132 transcripts per cell), and that the short transcript is most abundant in the early stationary phase cells (24 transcripts per cell).

Animals↗

PubMed1998

Body dissatisfaction and sexual victimization among college women.

109 white, never-married undergraduate women between the ages of 17 and 24 years completed both a Body Dissatisfaction scale and a sexual coercion scale. Analysis showed no over-all correlation between body-dissatisfaction scores and sexual victimization scores. Since the correlations were as anticipated, the lack of significance may reflect the small group sizes. Women with higher and those with lower body-dissatisfaction scores were not different in mean sexual victimization scores, suggesting that women's body dissatisfaction may not be associated with risk for sexual victimization. There was a significantly higher mean body-dissatisfaction score for women who reported that they had experienced sexual victimization in dating relationships than for women who reported having had no such experience. Perhaps experiencing sexual victimization has a negative effect on how women view their bodies. This study suggests a complex link between sexual victimization and body dissatisfaction in women.

Adolescent↗

PubMed1998

Short report: detection of Leishmaniavirus in human biopsy samples of leishmaniasis from Peru.

Leishmaniavirus is a double-stranded RNA virus that persistently infects some strains of the protozoan parasite Leishmania. There is considerable interest in the possibility that the presence of this virus alters parasite phenotype and may affect disease pathogenesis. If so, the virus marker could provide a valuable prognostic indicator for human leishmaniasis, particularly in those cases caused by New World parasite strains. The virus has been detected in cultured L. braziliensis, L. b. guyanensis, and L. major. To date there has been no information as to the extent of infection in samples prior to culturing in the laboratory. This study demonstrates, through the reverse transcription-polymerase chain reaction, that Leishmaniavirus exists in human biopsy samples of leishmaniasis prior to manipulation in culture.

Animals↗

PubMed1997

Cleavage site mapping and substrate-specificity of Leishmaniavirus 2-1 capsid endoribonuclease activity.

The Leishmaniavirus capsid protein possesses an RNA endoribonuclease activity that cleaves viral positive-sense RNA at a specific, single site within the 5' untranslated region. The site of cleavage in LRV1-4 RNA was previously mapped to nucleotide 320 of the LRV1-4 genome. Here we show that an LRV2-1-derived substrate RNA transcript is also cleaved at a single site in an in vitro cleavage assay with LRV2-1 virions. Precise RNA cleavage site mapping in this divergent Old World virus, LRV2-1, confirms that cleavage is occurring within a region of homology to the LRV1 isolates. Substrate RNA transcripts possessing viral sequences from LRV1-4 or LRV2-1 genomes were assayed for susceptibility to cleavage by the cognate and noncognate capsid endoribonucleases to determine the level of substrate specificity.

Binding Sites↗

PubMed1997

Specific in vitro cleavage of a Leishmania virus capsid-RNA-dependent RNA polymerase polyprotein by a host cysteine-like protease.

Antibodies raised against baculovirus-expressed RNA-dependent RNA polymerase (RDRP) recognized a 95-kDa antigen and two smaller proteins in sucrose-purified Leishmania virus particles isolated from infected parasites. The 95-kDa antigen is similar in size to one predicted by translation of the RDRP open reading frame (ORF) alone. In an effort to reconcile in vitro observations of translational frameshifting on Leishmania RNA virus 1-4 transcripts, we have developed an in vitro cleavage assay system to explore the possibility that the fusion polyprotein is proteolytically processed. We show that coincubation a synthetic Cap-Pol fusion protein with lysates from Leishmania parasites yields major cleavage products similar in size to those encoded by the individual capsid and RDRP genes as well as the antigens detected in vivo. The major 82- and 95-kDa major cleavage products are specifically immunoprecipitated by capsid- or polymerase-specific antibodies, respectively, showing that cleavage occurs at or near the junction of the two functional domains. Protease inhibitor studies suggest that a cysteine-like protease is responsible for cleavage in the in vitro assay system developed here. From these results, we suggest that failure to detect a capsid-polymerase fusion protein produced by translational frameshifting in vivo may be due to specific proteolytic processing.

Animals↗

PubMed1997

Hygromycin B resistance mediates elimination of Leishmania virus from persistently infected parasites.

A series of pX63-HYG derivatives encoding Leishmania RNA virus 1-4 (LRV1-4) sequences were electroporated into cells of Leishmania strain M4147, a virus-infected strain of L. guyanensis. After 6 weeks of drug selection (hygromycin B), transfected parasites lacked detectable quantities of viral genomic double-stranded RNA, viral capsid protein, and RNA-dependent RNA polymerase (RDRP) activity. Evidence of viral infection was not recovered upon removal of the drug. While viral RNA transcripts were produced from electroporated expression vectors, as determined by reverse transcription-PCR, viral antigens were not detected, suggesting that the antiviral effects of hygromycin B are mediated through translation inhibition. A short-term selection study suggests that the LRV1-4 elimination may not only be a function of hygromycin B as a protein synthesis inhibitor but also possibly related to the mechanism of hygromycin B resistance in Leishmania strains.

Animals↗

PubMed1996

Identification of a ribosomal frameshift in Leishmania RNA virus 1-4.

Double-stranded Leishmania RNA virus 1-4 (LRV 1-4) has at least four open reading frames (ORFs). The two small ORFs located near its 5' terminus, ORF1 and ORFx, could encode 34- and 60-amino acid polypeptides, respectively. ORF2 encodes an 82-kDa major capsid protein, and ORF3 encodes a 98-kDa polypeptide which contains the consensus sequence for RNA-dependent RNA polymerases of plus-strand and double-stranded RNA viruses. The complete sequence of LRV 1-4 shows that ORF2 and ORF3 overlap by 71 nucleotides, and that ORF3 lacks a potential translation initiation site, suggesting that the viral polymerase may be synthesized as a 180-kDa fusion protein with the virus capsid. In this report, we present evidence for the synthesis of a fusion protein through a ribosomal frameshift. In vitro-translation experiments and immunostudies involving antiserum against the viral capsid protein demonstrated that the overlapping 71 nucleotides of ORF2 and ORF3 are contained in a region which promotes translational frameshifting. Computer analysis of the putative frameshift region revealed a potential pseudoknot structure located within the overlapping 71 nucleotide sequence.

Amino Acid Sequence↗

PubMed1995

Single-site cleavage in the 5'-untranslated region of Leishmaniavirus RNA is mediated by the viral capsid protein.

Leishmaniavirus (LRV) is a double-stranded RNA virus that persistently infects the protozoan parasite Leishmania. LRV produces a short RNA transcript, corresponding to the 5' end of positive-sense viral RNA, both in vivo and in in vitro polymerase assays. The short transcript is generated by a single site-specific cleavage event in the 5' untranslated region of the 5.3-kb genome. This cleavage event can be reproduced in vitro with purified viral particles and a substrate RNA transcript possessing the viral cleavage site. A region of nucleotides required for cleavage was identified by analyzing the cleavage sites yielding the short transcripts of various LRV isolates. A 6-nt deletion at this cleavage site completely abolished RNA processing. In an in vitro cleavage assay, baculovirus-expressed capsid protein possessed an endonuclease activity identical to that of native virions, showing that the viral capsid protein is the RNA endonuclease. Identification of the LRV capsid protein as an RNA endonuclease is unprecedented among known viral capsid proteins.

Animals↗

PubMed1995

The complete sequence of Leishmania RNA virus LRV2-1, a virus of an Old World parasite strain.

A complete cDNA sequence is reported for LRV2-1, the first Leishmania RNA virus known to infect an Old World parasite, Leishmania major. Sequence analyses show that LRV2-1 differs significantly from members of the LRV1 genus which infect New World parasites. The data support a view that transmission of LRV is strictly vertical and suggest that LRV predate the divergence of Old and New World parasites. As a consequence of this divergence, conserved features can be identified for the first time in Leishmaniavirus proteins. A finding that the virus capsid and polymerase genes do not overlap is unique among the known Totiviridae and infers that a gag-pol fusion protein cannot be produced simply via tRNA slippage in LRV2-1.

Amino Acid Sequence↗

PubMed1995

The short transcript of Leishmania RNA virus is generated by RNA cleavage.

Leishmania RNA virus 1 produces a short viral RNA transcript corresponding to the 5' end of positive-sense single-stranded RNAs both in virally infected cells and in in vitro polymerase assays. We hypothesized that this short transcript was generated via cleavage of full-length positive-sense single-stranded RNA. A putative cleavage site was mapped by primer extension analysis to nucleotide 320 of the viral genome. To address the hypothesis that the short transcript is generated via cleavage at this site, two substrate RNAs that possessed viral sequence encompassing the putative cleavage site were created. When incubated with sucrose-purified viral particles, these substrate RNAs were site-specifically cleaved. The cleavage site of the in vitro-processed RNAs also mapped to viral nucleotide 320. The short-transcript-generating activity could be specifically abolished by proteinase K treatment of sucrose-purified viral particles and high concentrations of EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid], suggesting that the activity requires a proteinaceous factor and possibly intact viral particles. The cleavage activity is directly associated with short-transcript-generating activity, since only viral particle preparations which were capable of generating the short transcript in polymerase assays were also active in the cleavage assay. Furthermore, the short-transcript-generating activity is independent of the viral polymerase's transcriptase and replicase activities. We present a working model whereby cleavage of Leishmaniavirus RNA transcripts functions in the maintenance of a low-level persistent infection.

Animals↗

PubMed1995

Endogenous adenosine modulates alpha 2- but not alpha 1-adrenergic constriction of coronary arterioles.

In the coronary circulation alpha-adrenergic constriction competes with metabolic vasodilation. Because adenosine is produced by the working myocardium and metabolic stimulation results in arteriolar dilation, we tested the hypothesis that coronary arteriolar alpha-adrenergic constriction is attenuated by the endogenous production of adenosine. To test this hypothesis, using fluorescence microscopy during stroboscopic epi-illumination of the epicardial microvasculature, we measured the diameter of coronary arterioles in anesthetized open-chest dogs. Measurements were made in the presence of beta-blockade during selective alpha 1- or alpha 2-adrenoceptor activation (phenylephrine or B-HT-933, respectively) before and in the presence of the nonselective adenosine receptor antagonist 8-p-sulfophenyltheophylline (8-pSPT) and expressed as a percent change in microvascular diameter relative to baseline. alpha 1-Activation produced constriction of coronary arterioles under control conditions, which was not augmented after adenosine antagonism (-12 +/- 2 vs. -7 +/- 3%). In contrast, alpha 2-activation did not constrict coronary arterioles under control conditions; however, blockade of adenosine receptors unmasked a significant constriction (0 +/- 2 vs. -7 +/- 2%, P < 0.05). Also adenosine antagonism did not significantly alter the baseline diameter of coronary arterioles. These results demonstrate that endogenously produced adenosine modulates alpha 2-adrenergic constriction of coronary arterioles but not alpha 1-adrenergic constriction, and therefore we speculate that the competition between alpha-adrenergic constriction and metabolic vasodilation is mediated by the alpha 1-adrenoceptor.

Adenosine↗