Biomedical subjects

S E Johnson

Publications and source records attributed to S E Johnson.

At least 19 recordsLinked to original sources

PubMed2004

A murine model for the study of immune memory in response to pneumococcal conjugate vaccination.

We developed a murine model for assessment of immunological memory and antibody-induced protection to nasopharyngeal (NP) challenges. BALB/c female mice (n = 10 mice per study parameter) were immunized with two priming doses of the licensed 7-valent pneumococcal (Pnc) conjugate vaccine and immune responses [antibody immunoglobulin G (IgG) levels, avidity and opsonophagocytic activity] were monitored for 26 weeks until IgG levels decreased to nearly baseline. A booster dose of either 2 microg conjugate or 5 microg polysaccharide vaccine was given at week 26. The ability of these two treatments to recall immune memory established by the conjugate vaccine was determined for types 4 and 14 for up to 63 days post-booster. The ability of challenge with pneumococcal type 14 to recall the immune response was also evaluated, as well as, the number of antibody secreting cells (ASC) specific to polysaccharide (Ps) 4, 6B, and 14. A higher dose of conjugate vaccine (2 microg) was necessary to elicit a significant increase in IgG levels after priming with one dose. Priming with lower doses (0.5 and 1.0 microg) only elicited modest increases in IgG levels. Recall of the immune response was found with either conjugate or Ps vaccines. NP challenge with type 14 at week 26 did not recall the immune response, although reduction in NP Pnc load was seen post-primary immunization at 5, 10 and 26 weeks. ASCs were detected in response to either conjugate or Ps booster doses. This model allows for the screening and determination of potential alternative vaccination regimens and the study of immunological markers of memory following Pnc vaccination.

Animals↗

PubMed2001

Isolation and characterization of chicken TaxREB107, a putative DNA binding protein abundantly expressed in muscle.

Myogenic regulatory factors (MRFs) are vital transcription factors that act at multiple points during development to establish the skeletal muscle phenotype. This class of muscle-restricted, basic helix-loop-helix (bHLH) proteins acts in concert with additional transcriptional modulators to precisely control muscle gene expression. We have isolated the chicken homologue of Tax responsive element binding protein 107 (TaxREB107). The cDNA is 83% homologous at the amino acid level to human and mouse TaxREB107 and contains a centrally located leucine zipper motif. Northern analysis demonstrated that the gene is expressed in multiple tissues including skeletal muscle. Immunofluorescent staining revealed that the cTaxREB107 protein is located in both the nuclear and cytoplasmic compartments. Distinct localization to the nucleoli supports the evidence that TaxREB107 is a ribosomal protein. Because TaxREB proteins also are implicated in transcriptional regulation, we overexpressed cTaxREB107 in embryonic myoblasts. cTaxREB107 increased troponin I reporter gene activity as well as MRF-directed transcription from a multimerized skeletal muscle E-box reporter gene (4Rtk-luc). However, cotransfection of expression plasmids coding for MyoD and cTaxREB107 did not produce an increase in 4Rtk-luc suggesting that cTaxREB107 enhances myogenic gene transcription through a means independent of a physical association with MyoD. In conclusion, our results define a role for cTaxREB107 during avian myogenesis as a positive modulator of skeletal muscle gene expression.

Amino Acid Sequence↗

PubMed2001

Characterization of a dominant inhibitory E47 protein that suppresses C2C12 myogenesis.

Skeletal muscle formation is controlled through the coordinated actions of the muscle regulatory factors (MRFs). The activities of these basic helix-loop-helix proteins is mediated in part through heterodimer formation with a family of ubiquitous bHLH proteins, referred to as E-proteins. The primary E-protein in skeletal muscle is the E2A splice variant, E47. To further address the role of E47 during skeletal myogenesis, we created a chimeric E47 repressor protein by replacing the transcriptional activation domain with the Drosophila Engrailed transcriptional repressor domain. The dominant inhibitory E-protein (EnDeltaE47) formed homodimers capable of binding DNA and abolished E47-directed gene transcription. Stable expression of EnDeltaE47 in mouse C2C12 myoblasts effectively blocked the cells' ability to differentiate into mature myofibers. Closer examination of the molecular basis for the inhibition of myogenesis revealed that EnDeltaE47 preferentially forms heterodimers with myogenin. Interestingly, the chimeric repressor did not form DNA-binding heterodimers with MyoD in C2C12 myocytes. The failure to detect MyoD:EnDeltaE47 heterodimers in myoblasts was not due to protein conformational defects as both wild-type E47 and EnDeltaE47 readily formed DNA binding complexes with MyoD in vitro. These results indicate that E47 plays a crucial role in C2C12 myogenesis by serving as the preferred heterodimer partner of the myogenin protein.

Animals↗

PubMed2001

Idiopathic noncirrhotic portal hypertension in dogs: 33 cases (1982-1998).

OBJECTIVE: To describe clinical signs, diagnostic findings, and outcome in dogs with idiopathic intrahepatic portal hypertension. DESIGN: Retrospective study. ANIMALS: 33 dogs. PROCEDURE: Medical records of dogs with portal hypertension of intra-abdominal origin were reviewed. Dogs with intra-abdominal portal hypertension of vascular causes or with hepatic histopathologic changes consistent with severe diffuse hepatobiliary disease were excluded. History and results of physical examination, clinicopathologic tests, diagnostic imaging studies, histologic examination, and treatment were summarized. Outcome was determined in 26 dogs. RESULTS: Dogs were referred most often because of ascites, intermittent vomiting or diarrhea, and polydipsia of several months' duration. Microcytosis, high serum alkaline phosphatase and alanine transaminase activities, hepatic dysfunction, urine specific gravity < or = 1.021, and abdominal transudate were the predominant clinicopathologic features. Microhepatia, abdominal effusion, and multiple anomalous venous anastomoses were the major findings of diagnostic imaging. Hepatic histopathologic changes were consistent with idiopathic noncirrhotic portal hypertension and were indistinguishable from those of dogs with surgically created portocaval anastomosis. Outcome was determined for 19 dogs released from hospital; 13 dogs remained healthy with mostly palliative treatment for periods of 5 months to 9 years. CONCLUSIONS AND CLINICAL RELEVANCE: The clinical signs, clinicopathologic test results, portal pressure, and gross appearance of the liver of dogs with idiopathic noncirrhotic portal hypertension may be identical to those of dogs with cirrhosis; therefore liver biopsy is crucial. Because the prognosis for idiopathic noncirrhotic portal hypertension is generally favorable, owners of affected dogs should be discouraged from choosing euthanasia.

Alanine Transaminase↗

PubMed2001

Sensation seeking and hormones in men and women: exploring the link.

Risky behaviors (e.g., binge drinking, drunk driving, risky sex) are increasing among U.S. college students, and the personality trait of sensation seeking provides a potential link between such norm-breaking behaviors and biological processes. We examined the relationship between sensation-seeking behaviors and two hormones, testosterone and cortisol, in male and female college students. Hormone levels were hypothesized to contribute to the variability of individual scores on Zuckerman's Sensation-Seeking Scale. As expected, males scored higher on the scale than females, but the data failed to support the generally accepted positive relationship between testosterone and sensation seeking for either sex. Instead, our results support the existence of a significant inverse relationship between cortisol and sensation seeking in men, but not in women, even after adjustment for testosterone levels and age. Our study contributes to the current literature by (a) supporting the association between risky behavior and a hormone other than testosterone, (b) being the first to examine the association between cortisol and sensation seeking in women, and (c) identifying a possible effect of gender on the association between hormones and sensation-seeking behaviors. Gendered social norms and expectations are likely to be partly responsible for this effect. Theory-guided interdisciplinary research is needed to improve understanding of the biological influences on human behavior, and special attention must be paid to social context, women's perceptions of their expected behavior, and gendered socialization regarding norm-breaking or risky behaviors, which may obscure biological links to female behavior.

Adult↗

PubMed2001

Physical mapping of CSF2RA, ANT3 and STS on the pseudoautosomal region of bovine chromosome X.

The pseudoautosomal region (PAR) of bovine chromosome X (BTA X) has a particularly low representation of genes and markers, making comparative gene mapping in this region difficult. We describe the localization of three genes, colony-stimulating factor 2 receptor alpha (CSF2RA), ADP/ATP translocase 3 (ANT3) and steroid sulphatase (STS) on PAR of BTA X using a 5000 rad whole-genome radiation hybrid panel. The relationship of these genes to a number of previously mapped simple sequence repeat (microsatellite) markers is determined by physical and radiation hybrid mapping methods. The resulting radiation hybrid map resolves a discrepancy between the two major bovine linkage maps in the PAR of BTA X.

Animals↗

PubMed2000

Activated Raf inhibits myogenesis through a mechanism independent of activator protein 1-mediated myoblast transformation.

Skeletal myogenesis is acutely affected by growth factors and subsequent activation of their respective intracellular signaling cascades. Components of the mitogenic Ras/Raf/mitogen-activated protein kinase (MAPK) signaling module are potent inhibitors of myoblast differentiation. However, the means by which these kinases prevent myocyte formation and activation of the muscle gene program is unknown. Activator protein 1 (AP-1) is a transcriptional regulator the actions of which are up-regulated by signaling events, including elevated MAPK. Because activated Raf inhibits avian myogenesis in a MAPK-dependent fashion, we investigated the role of AP-1 as a mediator of the Raf-imposed block to myogenesis. Avian myoblasts overexpressing activated Raf contain elevated levels of AP-1 DNA binding and transcriptional activity. Introduction of an AP-1 dominant inhibitory protein (AFOS) into Raf-expressing myoblasts prevented acquisition of a transformed morphology. Interestingly, these cells remained differentiation-defective. Myogenic cells cotransduced with RCAS(A)-Raf BXB and RCAS(B)-AFOS remained mononuclear and myosin-negative and did not activate significantly muscle-specific reporter genes. These results argue that Raf inhibits muscle differentiation independent of AP-1-mediated cell transformation. Our results provide evidence for AP-1 as a critical component of the transforming capacity of activated Raf and evidence that AP-1 is not involved in the myogenic inhibitory effects of the kinase.

Animals↗

PubMed2000

Competition among Streptococcus pneumoniae for intranasal colonization in a mouse model.

Widespread use of conjugate vaccines against Streptococcus pneumoniae, by reducing carriage of S. pneumoniae serotypes included in the vaccine, may result in an increase in nasopharyngeal carriage of - and disease from - nonvaccine serotypes of the same species. Mathematical models predict that the extent of such replacement will depend positively on the degree to which carriage of vaccine-type S. pneumoniae inhibits acquisition of nonvaccine-type pneumococci, and may depend negatively on the inhibition of vaccine-type pneumococci by nonvaccine-type pneumococci. We used a mouse model of intranasal carriage of pneumococci to test whether such inhibition occurs between different pneumococcal strains. Mice carrying a streptomycin-resistant derivative of S. pneumoniae BG9163 (serotype 6B) as a resident strain showed reduced levels of colonization when challenged intranasally by optochin-resistant derivatives of the same strain and of a serotype 23F pneumococcus, BG8826. Inhibition could be overcome by increasing the dose of the challenge strain. Carriage of optochin-resistant BG9163 did not inhibit acquisition of the streptomycin-resistant variant. Colonization by a challenge strain did not significantly affect the level of colonization with the resident strain. These results provide evidence that is consistent with several hitherto untested assumptions of mathematical models of serotype replacement and suggest that a biological mechanism exists that could account for serotype replacement that is observed in clinical trials. The findings provide a basis for further studies of in vivo interactions between strains of S. pneumoniae.

Animals↗

PubMed2000

Purification and characterization of Streptococcus pneumoniae palmitoylated pneumococcal surface adhesin A expressed in Escherichia coli.

All Streptococcus pneumoniae isolates tested to date express a species-common lipoprotein designated as pneumococcal surface adhesin A (PsaA). This protein is cell-associated, hydrophobic, immunogenic, and genetically conserved. It is currently under investigation as a potential component in third-generation pneumococcal vaccine formulations. To overcome the problem of low-level expression of native hydrophobic PsaA in S. pneumoniae, and also of the recombinant PsaA (rPsaA) in Escherichia coli, we generated a stable E. coli construct expressing functional palmitoylated rPsaA ( approximately 10 mg/l of fermentation culture) using Borrelia burgdorferi outer surface protein A (OspA, a hydrophobic lipoprotein) signal peptide. By Western blot analysis, the chimeric rPsaA ( approximately 34 kDa) was detected in the cell lysate using anti-PsaA antibodies. It was partially purified by extracting the cell pellet with PBS/Triton X(R)-114 buffers, followed by anion exchange filter chromatography. A trypsin digestion profile of rPsaA closely resembled that of the native protein, as revealed by SDS-PAGE/silver staining. Lipidation of rPsaA was confirmed by labeling recombinant E. coli cells with [(3)H] palmitic acid and analyzing the labeled E. coli cells by Western blotting coupled with autoradiography. Further, analysis of purified rPsaA by mass spectrometry (MALDI-TOF) revealed a heterogenous spectrum with a major peak (M+H)(+1) of mass 33,384 Da (theoretical mass of palmitoylated rPsaA=33,361 Da). Purified rPsaA was immunogenic in CBA/NCAHN-XID female mice following intranasal immunization with or without adjuvant, as determined by measurement of anti-PsaA serum IgG levels. These anti-PsaA antibodies reacted with both native and rPsaA polypeptides. Our data strongly suggest that E. coli-expressed rPsaA is palmitoylated and closely resembles the native protein in structure and immunogenicity. It was also observed to elicit measurable protection against nasopharyngeal carriage with S. pneumoniae.

Adhesins, Bacterial↗

PubMed2000

FGF5 stimulates expansion of connective tissue fibroblasts and inhibits skeletal muscle development in the limb.

FGF5 is expressed in the mesenchyme and skeletal muscle of developing and adult mouse limbs. However, the function of FGF5 during development of the limb and limb musculature is unknown. To elucidate the inherent participation of FGF5 during limb organogenesis, a retroviral delivery system (RCAS) was used to overexpress human FGF5 throughout developing hind limb of chicken embryos. Misexpression of the soluble growth factor severely inhibited the formation of mature myocytes. Limbs infected with RCAS-FGF5 contained smaller presumptive muscle masses as evidenced by a decrease in MyoD and myosin heavy chain expressing cells. In contrast, ectopic expression of FGF5 significantly stimulated proliferation and expansion of the tenascin-expressing, connective-tissue fibroblast lineage throughout the developing limb. Histological analysis demonstrated that the increase in tenascin immunostaining surrounding the femur, ileum, and pubis in the FGF5 infected limbs corresponded to the fibroblasts forming the stacked-cell perichondrium. Furthermore, pulse labeling experiments with the thymidine analog, BrdU, revealed that the increased size of the perichondrium was attributable to enhanced cell proliferation. These results support a model whereby FGF5 acts as a mitogen to stimulate the proliferation of mesenchymal fibroblasts that contribute to the formation of connective tissues such as the perichondrium, and inhibits the development of differentiated skeletal muscle. These results also contend that FGF5 is a candidate mediator of the exclusive spatial patterning of the hind limb connective tissue and skeletal muscle.

Animals↗

PubMed2000

Expression and immune recognition of SV40 Tag in transgenic mice that develop metastatic osteosarcomas.

Mature adult mice of the C57BL/6-TgN(Amy1TAg)501Knw transgenic mouse lineage, 501, containing a liver alpha-amylase promoted-SV40 Tag hybrid gene, routinely develop SV40 Tag-induced metastatic osteosarcomas. This form of alpha-amylase was known to be expressed in the liver, salivary glands, pancreas, and fat. Cells in the normal rib adjacent to the periosteum also express alpha-amylase suggesting that transgene expression is correctly targeted to generate osteosarcomas. 501 mice express SV40 Tag in the salivary glands but do not develop abnormalities in these organs by the time of their death from SV40-induced osteosarcomas. Mice of the C57BL/6 strain make a strong and effective anti-tumor immune response to SV40 Tag immunization. However, immunization of 501 mice with SV40 Tag early in life does not alter or prevent SV40 Tag-induced osteosarcomagenesis. 501 mice mount a significantly less effective cytotoxic T-lymphocyte response following SV40 Tag immunization while 501 osteosarcoma-derived cells are fully susceptible to SV40 Tag-specific T-cell lysis. This suggests that partial tolerance, not loss of antigen presentation by tumor cells, characterizes this mouse model of endogenous bone tumor development. To determine whether the immune recognition of endogenous SV40 Tag could influence tumorigenesis, the metastatic potential and time of death from tumor was investigated in CD4-null mutant 501 mice and beta-2 microglobulin-null mutant 501 mice. The size and number of metastases in these strains and longevity of these strains varied. We suggest that components of both the innate and adaptive immune response control tumor appearance and progression.

Animals↗

PubMed2000

Teaching sedation and analgesia with simulation.

OBJECTIVE: This study reports on the efficacy of using the anesthesia simulator to teach sedation and analgesia to nurses. This provision of sedation and analgesia to a patient is accomplished with the goal of maintaining the ability of the patient to respond purposefully to auditory or tactile stimuli. METHODS: Nurses working in areas of the hospital where conscious sedation is performed were the participants in this sedation and analgesia training course. Prior to the training session, the participants read the American Society of Anesthesiology Practice guidelines for sedation and analgesia by non-anesthesiologists. At the time of the training session, each participant completed a written pretest, had an introduction to sedation and analgesia with four clinical crisis teaching scenarios using the anesthesia simulator, a practical exam using the simulator, and a written post-test. Each participant was also given the opportunity to complete an evaluation of the session. RESULTS: Twenty nurses completed the training session. The written tests had a maximum possible score of 30. Mean score on the written pretest was 22.9 +/- 3.54, and mean score on the written post-test was 26.0 +/- 4.24 (p < 0.001). Seventeen of the twenty subjects scored higher on the post-test. Mean practical exam score was 5.5 of a possible 6.0. Mean participant rating of the education session was 3.75 (1 = poor, 4 = excellent). All but one participant rated the length of the training session as "about right." CONCLUSIONS: The anesthesia simulator provides an excellent tool for teaching conscious sedation skills to hospital nurses. The participants' test performance improved following the session, and they also rated the educational experience as excellent.

Analgesia↗

PubMed2000

Prediction of neurologic morbidity in extremely low birth weight infants.

OBJECTIVE: (1) Identify major determinants of adverse neurodevelopmental outcome in extremely low birth weight (ELBW) infants. (2) Compare neural networks and regression analysis in the prediction of major handicaps and Bayley scores (MDI and PDI) in individual ELBW neonates followed to 18 months. STUDY DESIGN: Retrospective cohort study of regional tertiary care NICU database. A database with 21 selected variables was divided into training (n = 144) and test sets (n = 74). The training set was used to train a neural network and develop regression equations to predict outcomes in the test set. RESULTS: Determinants (descending order of contribution to variance): Major handicap: intraventricular hemorrhage (IVH) grade, necrotizing enterocolitis > or = stage II, black race, and no chorioamnionitis; low MDI: IVH grade, plurality, bronchopulmonary dysplasia (BPD), lower maternal grade, and no chorioamnionitis; low PDI: IVH grade, BPD, periventricular leukomalacia, lower maternal grade, and no chorioamnionitis. Regression techniques and neural networks were comparable and had relatively low sensitivity and correlation with adverse outcomes. CONCLUSION: Much of the variance in ELBW neurologic outcome cannot be explained by either regression analysis or neural network approaches.

Brain Diseases↗

PubMed2000

Insect chitin synthase cDNA sequence, gene organization and expression.

Chitin is a major component of the cuticle of arthropods. However, the synthesis of chitin is poorly understood. Feeding larvae of the insect Lucilia cuprina on the fungal chitin synthase competitive inhibitor, nikkomycin Z resulted in strong concentration-dependent mortality of the larvae (LD50 = 280 nM). This result demonstrates that chitin is an essential component of this insect. The complete cDNA and deduced amino-acid sequences of the first arthropod chitin synthase-like protein, LcCS-1, from the larvae of the insect L. cuprina have been determined. The cDNA sequence is 5757 bp in length and codes for a large complex protein containing 1592 amino acids (Mr = 180 717). Analysis of the whole protein sequence reveals low, but significant, similarity to yeast chitin synthases with stronger areas of conservation centred on local regions implicated in the active sites of the yeast enzymes. Strikingly, LcCS-1 contains 15-18 potential transmembrane segments, indicating that the protein is an integral membrane protein. Two alternative topographical models of LcCS-1 are described, which involve its association with either the plasma membrane or the membrane of intracellular vesicles. LcCS-1 mRNA is produced in all life stages of the insect with expression in the larval stage limited to the integument and trachea. In a third instar larva the mRNA was localized to a single layer of epidermal cells immediately underlying the procuticle region of the integument. cDNA or genomic sequences that are highly related to fragments of LcCS-1 were demonstrated in three insect orders, one arachnid and Caenorhabditis elegans, thereby attesting to the importance of this enzyme in these chitin-producing organisms. Bioinformatics has been used to deduce the gene sequence and organization of the highly homologous Drosophila melanogaster orthologue of LcCS-1, DmCS-1.

Amino Acid Sequence↗

PubMed2000

Selection of an immunogenic and protective epitope of the PsaA protein of Streptococcus pneumoniae using a phage display library.

Streptococcus pneumoniae is an important pathogen that causes disease in young and elderly individuals. The currently available polysaccharide vaccines have limited efficacy in those age groups most susceptible to pneumococcal infections. This study focuses on mapping the epitopes of a surface protein of S. pneumoniae by biopanning a 15 mer phage display library using 5 different monoclonal antibodies (MAbs) against the Pneumoccal surface adhesin A (PsaA). PsaA is a component of the bacterial cell wall that is highly species specific and is involved in bacterial adherence and virulence. Biopanning of the phage display library reveals three distinct epitopes on the PsaA protein. The sequence homology of these epitopes ranges from two to six amino acids when compared to the native PsaA protein type 2. Two of these epitopes have been evaluated for their immunogeneicity in mice. The peptide selected by the MAbs 8G12, 6F6, and 1B7 is referred to as the consensus peptide and is immunogenic in mice. Optimal anti-PsaA response is observed in mice immunized with 50microg of the consensus peptide complexed to proteosomes in 1:1 ratio. The anti-PsaA response is significantly lower than the response to the PsaA native protein. The peptide selected by monoclonal antibody 4E9 in its lipidated form is significantly protective in mice challenged with S. pneumoniae serotype 2 when compared to mice immunized with the native protein. These results show that the selected epitopes of PsaA protein are immunogenic and protective in mice. These epitopes need to be evaluated further as alternatives to currently available vaccines.

Adhesins, Bacterial↗

PubMed1999

Isolation of a cDNA encoding a putative cellulase in the red claw crayfish Cherax quadricarinatus.

Amino acid sequences of cellulases have been determined in insects, nematodes, plants, slime moulds and bacteria but not in crustaceans. However, cellulase activity has been demonstrated in the hepatopancreas of the red claw crayfish, Cherax quadricarinatus. In order to obtain information on the nature of this cellulase, a C. quadricarinatus hepatopancreas cDNA library was screened with a PCR product generated using degenerate oligonucleotide primers derived from conserved regions of known cellulases. Two identical 1.56kb cDNAs with sequence similarities to known cellulases, particularly the termite endoglucanases, were identified and sequenced. The clones contain the complete cDNA open reading frame for an endo-1, 4-beta-glucanase of 469 amino acids termed Cherax quadricarinatus endoglucanase (CqEG). The endogenous origin of the gene was confirmed by PCR amplification and sequencing of a 1012bp PCR product from genomic DNA. This fragment contains four exon sequences identical to the cDNA and is interrupted by three introns of 371, 102, 194bp respectively, with one intron exhibiting typical eukaryotic splice sites. The isolation of an endo-1,4-beta-glucanase encoding cDNA from the crayfish C. quadricarinatus provides the first endogenous cellulase sequence in a crustacean species.

Amino Acid Sequence↗

PubMed1999

Activated Raf inhibits avian myogenesis through a MAPK-dependent mechanism.

Chronic overexpression of the oncogenic form of Ras is a potent inhibitor of skeletal myogenesis. However, the intracellular signaling pathways that mediate the repressive actions of Ras on myogenic differentiation have yet to be identified. We examined the role of Raf-mediated signaling as a modulator of avian myogenesis. Raf overexpression elicited pronounced effects on both myoblasts and mature myocytes. Most notably, the embryonic chick myoblasts overexpressing a constitutively active form of Raf (RCAS-Raf CAAX or RCAS-Raf BXB) fail to form the large multinucleated myofibers characteristic of myogenic cultures. While residual myofibers were apparent in the RCAS-Raf BXB and RCAS-Raf CAAX infected cultures, these fibers had an atrophic phenotype. The altered morphology is not a result of reinitiation of the myonuclei cell cycle nor is it due to apoptosis. Furthermore, the mononucleated myoblasts misexpressing Raf BXB are differentiation-defective due to overt MAPK activity. Supplementation of the culture media with the MAPK kinase (MEK) inhibitor, PD98059, caused a reversal of the phenotype and allowed the formation of multinucleated myofibers at levels comparable to controls. Our results indicate that the Raf/MEK/MAPK axis is intact in chick myoblasts and that persistent activation of this signaling cascade is inhibitory to myogenesis.

Animals↗

PubMed1999

Census of brown lemurs (Eulemur fulvus sspp.) in southeastern Madagascar: methods-testing and conservation implications.

Four forest areas were censused in southeastern Madagascar from June-August 1995 to estimate local population densities and habitat conditions for two threatened subspecies of brown lemur (Eulemur fulvus collaris and Eulemur fulvus albocollaris). Survey transects varied in length (1-3.5 km) and in surveillance frequency (three to seven times). Additional test surveys were conducted at the Parc National de Ranomafana to compare transect methods in an area with known population densities of Eulemur fulvus. Based on these tests, we demonstrate that the use of existing trails for transects can result in a close estimate of local population size, although more replications and longer transects increase precision. Results from regional surveys indicate considerably smaller population densities for E. f. albocollaris (0.086 animals/ha). In contrast, E. f. collaris densities were relatively high (.107 animals/ha) at Midongy-Sud. We also noted variation among sites in the density of lianas, which was positively correlated with local population density (a possible indication of habitat degradation). More generally, habitats in E. f. albocollaris's range suffered from fragmentation, reduction in forest area, logging, and potentially greater hunting pressure. Based on these results, it is apparent that more immediate steps are necessary to preserve E. f. albocollaris populations and habitats.

Animals↗