Derek Eberhart - Academia.edu (original) (raw)

Papers by Derek Eberhart

Research paper thumbnail of Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from <i>Cxcr6</i> Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Supplemental Figure S1 Total RNA samples (10 µg) extracted from adult cerebella were analyzed by ... more Supplemental Figure S1 Total RNA samples (10 µg) extracted from adult cerebella were analyzed by Northern blotting using radiolabeled Fyco1 and Gapdh probes. Genotype of each sample is indicated at the top of panels.

Research paper thumbnail of Assessing the Effects of the Naked Neck Gene on Chronic Heat Stress Resistance in Two Genetic Populations

Poultry Science, Aug 1, 1993

The effect of the naked neck (Na) gene on resistance to chronic heat stress was studied in the F2... more The effect of the naked neck (Na) gene on resistance to chronic heat stress was studied in the F2 generation of two populations of chickens genetically differing in growth. The Na gene was introduced into the Athens-Canadian randombred (ACRB), a small BW population, and into a large BW commercial broiler population. Naked neck and normally feathered birds were maintained in either a chronic heat stress (32 C) environment or a control (21 C) environment from 4 to 8 wk of age. Body weight, BW gain, feed consumption, and feed efficiency were calculated at 4, 6, and 8 wk. The 32 C environment significantly reduced gain and feed consumption at all age intervals in both populations. Feed conversion ratio (FCR) was lower in the 32 C environment in the ACRB population, but in the broiler population FCR either was the same or increased when compared with the 21 C environment. In the F2 generation of both populations the naked neck birds were significantly larger than the normally feathered birds although the two types segregated from the same parents. In the ACRB population the relative growth response was not significantly different between naked neck and normal feathered birds, indicating that the Na gene did not confer resistance to chronic heat stress in the small BW population. In the broiler population, an assessment of the relative growth response in the 32 C environment showed that the naked neck birds had a smaller reduction in BW gain and better feed efficiency than the normally feathered birds, indicating that in the large BW broiler population the Na gene did confer resistance to chronic heat stress.

Research paper thumbnail of Data from Medulloblastomas Derived from <i>Cxcr6</i> Mutant Mice Respond to Treatment with a Smoothened Inhibitor

The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in incre... more The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small-molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. [Cancer Res 2007;67(8):3871–7]

Research paper thumbnail of Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respon... more Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Research paper thumbnail of Data from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in incre... more The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that redu...

Research paper thumbnail of Pediatrics, and

Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unsta... more Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unstable expansion of a CGGrepeat within the FMR1 gene that results in the absence of the encoded protein. The fragile X mental retardation protein (FMRP) is a ribosome-associated RNA-binding protein of uncertain function that contains nuclear localization and export signals. We show here detailed cellular localization studies using both biochemical and immunocytochemical approaches. FMRP was highly expressed in neurons but not glia throughout the rat brain, as detected by light microscopy. Although certain structures, such as hippocampus, revealed a strong signal, the regional variation in staining intensity appeared to be related to neuron size and density. In human cell lines and mouse brain, FMRP co-fractionated primarily with polysomes and rough endoplasmic reticulum. Ultrastructural studies in rat brain revealed

Research paper thumbnail of For Patched 1

This article cites 27 articles, 11 of which you can access for free at:

Research paper thumbnail of Triagem De Tecnologias – Ferramenta Para a Construção e Gestão De Um Portfólio Tecnológico Robusto Em Inovação e Transferência De Tecnologia

Revista Gestão, Inovação e Tecnologias, 2013

The assessment of the technological potential, of the patentability and of the commercial potenti... more The assessment of the technological potential, of the patentability and of the commercial potential of inventions developed at universities is generally a very difficult process, often subject to errors in evaluation. The key problem faced by academic Technology Transfer Offices (TTO) is the very incipient stage of the technologies developed at universities, which are-routinely-closer to the level of a discovery than that of a finished product. Over the years, TTOs have developed tools that facilitate and bring a bit more precision to the assessment process. In Brazil, the development of such tool, suited to the local reality, was pioneered by INOVA-UniCamp. These tools are called "Triage Forms", two of which are introduced here to the Brazilian technology transfer community.

Research paper thumbnail of Métodos e ferramentas para avaliação de tecnologias em estágio embrionário

Capacite: Os Caminhos para a Inovação Tecnológica, 2014

Research paper thumbnail of Cytochrome P450 IIIA1 (P450p) requires cytochrome b5 and phospholipid with unsaturated fatty acids

Archives of Biochemistry and Biophysics, 1991

Research paper thumbnail of Variation in Body Temperature Response of Naked Neck and Normally Feathered Chickens to Heat Stress

Poultry Science, 1993

Abstract The effect of the naked neck (Na) gene on basal body temperature (BT) in a 21 C (control... more Abstract The effect of the naked neck (Na) gene on basal body temperature (BT) in a 21 C (control) or in a 32 C (chronic heat stress) environment and on change in BT when exposed to acute heat stress conditions (40.5 C) was examined in small and large BW populations. Birds grown at 32 C had a significantly lower basal BT, 8-wk BW, and a significantly smaller change in BT when exposed to 40.5 C than birds grown in 21 C. The small BW birds had a higher basal BT and a smaller change in BT when exposed to acute heat stress than large BW birds. There was no difference in basal BT between naked neck and normal birds in either the small BW (Athens-Canadian randombred) population, or in the large BW broiler population. The Na/Na birds in the small BW population had a significantly greater change in BT when exposed to acute heat stress than Na/na or na/na birds, but no differences were seen among genotypes in the large BW population. In the 21 C environment, naked neck birds seemed to have a greater change in BT than normal birds when exposed to 40.5 C. The difference in BT change was significant in the large BW population but was not significant in the small BW population. Few significant correlations were found between basal BT and 8-wk BW or between change in BT and 8-wk BW. A consistent negative correlation was found between basal BT and change in BT for both populations.

Research paper thumbnail of The fragile X mental retardation protein is a ribonucleoprotein containing both nuclear localization and nuclear export signals

Human Molecular Genetics, 1996

Fragile X syndrome is a frequent cause of mental retardation resulting from the absence of FMRP, ... more Fragile X syndrome is a frequent cause of mental retardation resulting from the absence of FMRP, the protein encoded by the FMR1 gene. FMRP is an RNA-binding protein of unknown function which is associated with ribosomes. To gain insight into FMRP function, we performed immunolocalization analysis of FMRP truncation and fusion constructs which revealed a nuclear localization signal (NLS) in the amino terminus of FMRP as well as a nuclear export signal (NES) encoded by exon 14. A 17 amino acid peptide containing the FMRP NES, which closely resembles the NES motifs recently described for HIV-1 Rev and PKI, is sufficient to direct nuclear export of a microinjected protein conjugate. Sucrose gradient analysis shows that FMRP ribosome association is RNA-dependent and FMRP is found in ribonucleoprotein (RNP) particles following EDTA treatment. These data are consistent with nascent FMRP entering the nucleus to assemble into mRNP particles prior to export back into the cytoplasm and suggests that fragile X syndrome may result from altered translation of transcripts which normally bind to FMRP.

Research paper thumbnail of Loss of p53 but not ARF Accelerates Medulloblastoma in Mice Heterozygous for patched1

Cancer Research, 2001

Brain malignancies represent the most common solid tumors in children, and they are responsible f... more Brain malignancies represent the most common solid tumors in children, and they are responsible for significant mortality and morbidity. The molecular basis of the most common malignant pediatric brain tumor, medulloblastoma, is poorly understood. Mutations in several genes including the human homologue of the Drosophila segment polarity gene, patched (PTCH), the adenomatous polyposis coli gene (APC), ␤-catenin, and p53 have been reported in subsets of hereditary and sporadic medulloblastoma. Inactivation of one Ptc allele in mice results in a 14% incidence of medulloblastoma. Here, we report a dramatic increase in the incidence (>95%) and accelerated development (prior to 12 weeks of age) of medulloblastoma in mice heterozygous for Ptc that lack p53. The acceleration of tumorigenesis in Ptc ؉/؊ mice is specific for loss of p53, because no change in tumor incidence was observed in Ptc ؉/؊ mice carrying a mutation in APC (Min ؉/؊) or in Ptc ؉/؊ mice deficient in p19 ARF. Thus, there is a specific interaction between p53 loss and heterozygosity of Ptc that results in medulloblastoma. This may be a consequence of increased genomic instability associated with loss of p53 function that may enhance the rate of acquisition of secondary mutations. Ptc ؉/؊ p53 ؊/؊ mice provide a useful model for investigation of the molecular bases of medulloblastoma and for evaluation of the efficacy of therapeutic intervention strategies in a spontaneously arising endogenous brain tumor.

Research paper thumbnail of Decreased Intraocular Pressure in Mice Following Either Pharmacological or Genetic Inhibition of ROCK

Journal of Ocular Pharmacology and Therapeutics the Official Journal of the Association For Ocular Pharmacology and Therapeutics, Jul 1, 2009

Goals of this study were to determine if pharmacological or genetic inhibition of Rho-associated ... more Goals of this study were to determine if pharmacological or genetic inhibition of Rho-associated coiled coil containing protein kinases (known as ROCK1 and ROCK2) alters intraocular pressure (IOP) in mice. Micro-cannulation of the anterior chamber was used to measure IOP in wild-type B6.129 hybrid mice following treatment with ROCK inhibitors Y-27632 or Y-39983. For comparative purposes, wild-type mice were also treated with timolol, acetazolamide, pilocarpine, or latanoprost. Mice deficient in either Rock1 or Rock2 were generated by homologous recombination or gene trapping, respectively, and their IOP was determined using identical methods employed in the pharmacology studies. Treatment of wild-type B6.129 hybrid mice with ROCK inhibitors (Y-27632 and Y-39983) resulted in significant reductions in IOP. The magnitude of IOP reduction observed with topical Y-39983 was comparable to timolol, and exceeded the IOP effects of latanoprost in this study. Pilocarpine had no discernible effect on IOP in mice. Moreover, mice deficient in either Rock1 or Rock2 exhibited a significant decrease in IOP compared to their B6.129 wild-type littermates. Pharmacological or genetic inhibition of ROCKs results in decreased IOP in mice. The magnitude of IOP reduction is significant as demonstrated with comparative pharmacology using agents that lower IOP in humans. These studies support the ROCK pathway as a therapeutic target for treating ocular hypertension.

Research paper thumbnail of The Normal patched Allele Is Expressed in Medulloblastomas from Mice with Heterozygous Germ-Line Mutation of patched1

Cancer Research, Apr 15, 2000

Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila seg... more Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila segment polarity gene, patched (ptc) and its ligand, sonic hedgehog (shh), contribute to tumor formation in several tissues. Recently, a subset of medulloblastoma, the most common malignant brain tumor in children, was found to contain somatic mutations in the human ptc gene. In addition, basal cell nevus syndrome (BCNS), or Gorlin syndrome, which is characterized by developmental anomalies and a predisposition to skin and nervous system malignancies, is associated with germ-line mutation of ptc. Targeted disruption of both alleles of ptc in mice results in embryonic lethality. However, ptc ؉/؊ mice survive and develop spontaneous cerebellar brain tumors, suggesting that ptc may function as a tumor suppressor gene. Therefore, we investigated ptc ؉/؊ mice as a model for human medulloblastoma. We report that 14% of ptc ؉/؊ mice develop central nervous system tumors in the posterior fossa by 10 months of age, with peak tumor incidence occurring between 16 and 24 weeks of age. The tumors exhibited several characteristics of human medulloblastoma, including expression of intermediate filament proteins specific for neurons and glia. Full-length ptc mRNA was present in all tumors analyzed, indicating that there was no loss of heterozygosity at the ptc locus. Nucleotide sequence of ptc mRNA from four tumors failed to identify any mutations. However, a comparison of the normal ptc sequence from C57BL/6 and 129Sv mice did reveal several polymorphisms. High levels of gli1 mRNA and protein were detected in the tumors, suggesting that the shh/ptc pathway was activated despite the persistence of ptc expression. These data indicate that haploinsufficiency of ptc is sufficient to promote oncogenesis in the central nervous system.

Research paper thumbnail of The molecular basis of fragile X syndrome

Cold Spring Harbor symposia on quantitative biology, 1996

Research paper thumbnail of Cysteine 64 of Ref-1 is not essential for redox regulation of AP-1 DNA binding

Molecular and cellular biology, 2003

Ref-1 participates in DNA repair as well as in redox regulation of transcription factor function.... more Ref-1 participates in DNA repair as well as in redox regulation of transcription factor function. The redox function of Ref-1 involves reduction of oxidized cysteine residues within the DNA binding domains of several transcription factors, including Fos and Jun. Reduction of these residues is required for DNA binding, providing a redox-dependent mechanism for regulation of target gene expression. Previous in vitro studies implicated cysteine 65 of human Ref-1 (cysteine 64 of mouse Ref-1) as the redox catalytic site. We analyzed the in vivo role of cysteine 64 in redox regulation of AP-1 activity by introducing a cysteine-to-alanine point mutation into the endogenous mouse Ref-1 gene (ref-1(C64A)). Unlike Ref-1 null mice, which die very early in embryonic development, homozygous ref-1(C64A) mice are viable, they survive to normal life expectancy, and they display no overt abnormal phenotype. Although Ref-1 provides the major AP-1-reducing activity in murine cells, ref-1(C64A) cells r...

Research paper thumbnail of Loss of p53 but not ARF accelerates medulloblastoma in mice heterozygous for patched

Cancer research, Jan 15, 2001

Brain malignancies represent the most common solid tumors in children, and they are responsible f... more Brain malignancies represent the most common solid tumors in children, and they are responsible for significant mortality and morbidity. The molecular basis of the most common malignant pediatric brain tumor, medulloblastoma, is poorly understood. Mutations in several genes including the human homologue of the Drosophila segment polarity gene, patched (PTCH), the adenomatous polyposis coli gene (APC), beta-catenin, and p53 have been reported in subsets of hereditary and sporadic medulloblastoma. Inactivation of one Ptc allele in mice results in a 14% incidence of medulloblastoma. Here, we report a dramatic increase in the incidence (>95%) and accelerated development (prior to 12 weeks of age) of medulloblastoma in mice heterozygous for Ptc that lack p53. The acceleration of tumorigenesis in Ptc+/- mice is specific for loss of p53, because no change in tumor incidence was observed in Ptc+/- mice carrying a mutation in APC (Min+/-) or in Ptc+/- mice deficient in p19ARF. Thus, there...

Research paper thumbnail of The normal patched allele is expressed in medulloblastomas from mice with heterozygous germ-line mutation of patched

Cancer research, Jan 15, 2000

Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila seg... more Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila segment polarity gene, patched (ptc) and its ligand, sonic hedgehog (shh), contribute to tumor formation in several tissues. Recently, a subset of medulloblastoma, the most common malignant brain tumor in children, was found to contain somatic mutations in the human ptc gene. In addition, basal cell nevus syndrome (BCNS), or Gorlin syndrome, which is characterized by developmental anomalies and a predisposition to skin and nervous system malignancies, is associated with germ-line mutation of ptc. Targeted disruption of both alleles of ptc in mice results in embryonic lethality. However, ptc+/- mice survive and develop spontaneous cerebellar brain tumors, suggesting that ptc may function as a tumor suppressor gene. Therefore, we investigated ptc+/-mice as a model for human medulloblastoma. We report that 14% of ptc+/- mice develop central nervous system tumors in the posterior fossa by 10 mo...

Research paper thumbnail of Fragile X mental retardation protein: nucleocytoplasmic shuttling and association with somatodendritic ribosomes

The Journal of neuroscience : the official journal of the Society for Neuroscience, 1997

Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unsta... more Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unstable expansion of a CGG-repeat within the FMR1 gene that results in the absence of the encoded protein. The fragile X mental retardation protein (FMRP) is a ribosome-associated RNA-binding protein of uncertain function that contains nuclear localization and export signals. We show here detailed cellular localization studies using both biochemical and immunocytochemical approaches. FMRP was highly expressed in neurons but not glia throughout the rat brain, as detected by light microscopy. Although certain structures, such as hippocampus, revealed a strong signal, the regional variation in staining intensity appeared to be related to neuron size and density. In human cell lines and mouse brain, FMRP co-fractionated primarily with polysomes and rough endoplasmic reticulum. Ultrastructural studies in rat brain revealed high levels of FMRP immunoreactivity in neuronal perikarya, where it is co...

Research paper thumbnail of Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from <i>Cxcr6</i> Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Supplemental Figure S1 Total RNA samples (10 µg) extracted from adult cerebella were analyzed by ... more Supplemental Figure S1 Total RNA samples (10 µg) extracted from adult cerebella were analyzed by Northern blotting using radiolabeled Fyco1 and Gapdh probes. Genotype of each sample is indicated at the top of panels.

Research paper thumbnail of Assessing the Effects of the Naked Neck Gene on Chronic Heat Stress Resistance in Two Genetic Populations

Poultry Science, Aug 1, 1993

The effect of the naked neck (Na) gene on resistance to chronic heat stress was studied in the F2... more The effect of the naked neck (Na) gene on resistance to chronic heat stress was studied in the F2 generation of two populations of chickens genetically differing in growth. The Na gene was introduced into the Athens-Canadian randombred (ACRB), a small BW population, and into a large BW commercial broiler population. Naked neck and normally feathered birds were maintained in either a chronic heat stress (32 C) environment or a control (21 C) environment from 4 to 8 wk of age. Body weight, BW gain, feed consumption, and feed efficiency were calculated at 4, 6, and 8 wk. The 32 C environment significantly reduced gain and feed consumption at all age intervals in both populations. Feed conversion ratio (FCR) was lower in the 32 C environment in the ACRB population, but in the broiler population FCR either was the same or increased when compared with the 21 C environment. In the F2 generation of both populations the naked neck birds were significantly larger than the normally feathered birds although the two types segregated from the same parents. In the ACRB population the relative growth response was not significantly different between naked neck and normal feathered birds, indicating that the Na gene did not confer resistance to chronic heat stress in the small BW population. In the broiler population, an assessment of the relative growth response in the 32 C environment showed that the naked neck birds had a smaller reduction in BW gain and better feed efficiency than the normally feathered birds, indicating that in the large BW broiler population the Na gene did confer resistance to chronic heat stress.

Research paper thumbnail of Data from Medulloblastomas Derived from <i>Cxcr6</i> Mutant Mice Respond to Treatment with a Smoothened Inhibitor

The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in incre... more The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small-molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. [Cancer Res 2007;67(8):3871–7]

Research paper thumbnail of Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respon... more Supplementary Figure 1 and Tables 1-2 from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

Research paper thumbnail of Data from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor

The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in incre... more The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that redu...

Research paper thumbnail of Pediatrics, and

Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unsta... more Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unstable expansion of a CGGrepeat within the FMR1 gene that results in the absence of the encoded protein. The fragile X mental retardation protein (FMRP) is a ribosome-associated RNA-binding protein of uncertain function that contains nuclear localization and export signals. We show here detailed cellular localization studies using both biochemical and immunocytochemical approaches. FMRP was highly expressed in neurons but not glia throughout the rat brain, as detected by light microscopy. Although certain structures, such as hippocampus, revealed a strong signal, the regional variation in staining intensity appeared to be related to neuron size and density. In human cell lines and mouse brain, FMRP co-fractionated primarily with polysomes and rough endoplasmic reticulum. Ultrastructural studies in rat brain revealed

Research paper thumbnail of For Patched 1

This article cites 27 articles, 11 of which you can access for free at:

Research paper thumbnail of Triagem De Tecnologias – Ferramenta Para a Construção e Gestão De Um Portfólio Tecnológico Robusto Em Inovação e Transferência De Tecnologia

Revista Gestão, Inovação e Tecnologias, 2013

The assessment of the technological potential, of the patentability and of the commercial potenti... more The assessment of the technological potential, of the patentability and of the commercial potential of inventions developed at universities is generally a very difficult process, often subject to errors in evaluation. The key problem faced by academic Technology Transfer Offices (TTO) is the very incipient stage of the technologies developed at universities, which are-routinely-closer to the level of a discovery than that of a finished product. Over the years, TTOs have developed tools that facilitate and bring a bit more precision to the assessment process. In Brazil, the development of such tool, suited to the local reality, was pioneered by INOVA-UniCamp. These tools are called "Triage Forms", two of which are introduced here to the Brazilian technology transfer community.

Research paper thumbnail of Métodos e ferramentas para avaliação de tecnologias em estágio embrionário

Capacite: Os Caminhos para a Inovação Tecnológica, 2014

Research paper thumbnail of Cytochrome P450 IIIA1 (P450p) requires cytochrome b5 and phospholipid with unsaturated fatty acids

Archives of Biochemistry and Biophysics, 1991

Research paper thumbnail of Variation in Body Temperature Response of Naked Neck and Normally Feathered Chickens to Heat Stress

Poultry Science, 1993

Abstract The effect of the naked neck (Na) gene on basal body temperature (BT) in a 21 C (control... more Abstract The effect of the naked neck (Na) gene on basal body temperature (BT) in a 21 C (control) or in a 32 C (chronic heat stress) environment and on change in BT when exposed to acute heat stress conditions (40.5 C) was examined in small and large BW populations. Birds grown at 32 C had a significantly lower basal BT, 8-wk BW, and a significantly smaller change in BT when exposed to 40.5 C than birds grown in 21 C. The small BW birds had a higher basal BT and a smaller change in BT when exposed to acute heat stress than large BW birds. There was no difference in basal BT between naked neck and normal birds in either the small BW (Athens-Canadian randombred) population, or in the large BW broiler population. The Na/Na birds in the small BW population had a significantly greater change in BT when exposed to acute heat stress than Na/na or na/na birds, but no differences were seen among genotypes in the large BW population. In the 21 C environment, naked neck birds seemed to have a greater change in BT than normal birds when exposed to 40.5 C. The difference in BT change was significant in the large BW population but was not significant in the small BW population. Few significant correlations were found between basal BT and 8-wk BW or between change in BT and 8-wk BW. A consistent negative correlation was found between basal BT and change in BT for both populations.

Research paper thumbnail of The fragile X mental retardation protein is a ribonucleoprotein containing both nuclear localization and nuclear export signals

Human Molecular Genetics, 1996

Fragile X syndrome is a frequent cause of mental retardation resulting from the absence of FMRP, ... more Fragile X syndrome is a frequent cause of mental retardation resulting from the absence of FMRP, the protein encoded by the FMR1 gene. FMRP is an RNA-binding protein of unknown function which is associated with ribosomes. To gain insight into FMRP function, we performed immunolocalization analysis of FMRP truncation and fusion constructs which revealed a nuclear localization signal (NLS) in the amino terminus of FMRP as well as a nuclear export signal (NES) encoded by exon 14. A 17 amino acid peptide containing the FMRP NES, which closely resembles the NES motifs recently described for HIV-1 Rev and PKI, is sufficient to direct nuclear export of a microinjected protein conjugate. Sucrose gradient analysis shows that FMRP ribosome association is RNA-dependent and FMRP is found in ribonucleoprotein (RNP) particles following EDTA treatment. These data are consistent with nascent FMRP entering the nucleus to assemble into mRNP particles prior to export back into the cytoplasm and suggests that fragile X syndrome may result from altered translation of transcripts which normally bind to FMRP.

Research paper thumbnail of Loss of p53 but not ARF Accelerates Medulloblastoma in Mice Heterozygous for patched1

Cancer Research, 2001

Brain malignancies represent the most common solid tumors in children, and they are responsible f... more Brain malignancies represent the most common solid tumors in children, and they are responsible for significant mortality and morbidity. The molecular basis of the most common malignant pediatric brain tumor, medulloblastoma, is poorly understood. Mutations in several genes including the human homologue of the Drosophila segment polarity gene, patched (PTCH), the adenomatous polyposis coli gene (APC), ␤-catenin, and p53 have been reported in subsets of hereditary and sporadic medulloblastoma. Inactivation of one Ptc allele in mice results in a 14% incidence of medulloblastoma. Here, we report a dramatic increase in the incidence (>95%) and accelerated development (prior to 12 weeks of age) of medulloblastoma in mice heterozygous for Ptc that lack p53. The acceleration of tumorigenesis in Ptc ؉/؊ mice is specific for loss of p53, because no change in tumor incidence was observed in Ptc ؉/؊ mice carrying a mutation in APC (Min ؉/؊) or in Ptc ؉/؊ mice deficient in p19 ARF. Thus, there is a specific interaction between p53 loss and heterozygosity of Ptc that results in medulloblastoma. This may be a consequence of increased genomic instability associated with loss of p53 function that may enhance the rate of acquisition of secondary mutations. Ptc ؉/؊ p53 ؊/؊ mice provide a useful model for investigation of the molecular bases of medulloblastoma and for evaluation of the efficacy of therapeutic intervention strategies in a spontaneously arising endogenous brain tumor.

Research paper thumbnail of Decreased Intraocular Pressure in Mice Following Either Pharmacological or Genetic Inhibition of ROCK

Journal of Ocular Pharmacology and Therapeutics the Official Journal of the Association For Ocular Pharmacology and Therapeutics, Jul 1, 2009

Goals of this study were to determine if pharmacological or genetic inhibition of Rho-associated ... more Goals of this study were to determine if pharmacological or genetic inhibition of Rho-associated coiled coil containing protein kinases (known as ROCK1 and ROCK2) alters intraocular pressure (IOP) in mice. Micro-cannulation of the anterior chamber was used to measure IOP in wild-type B6.129 hybrid mice following treatment with ROCK inhibitors Y-27632 or Y-39983. For comparative purposes, wild-type mice were also treated with timolol, acetazolamide, pilocarpine, or latanoprost. Mice deficient in either Rock1 or Rock2 were generated by homologous recombination or gene trapping, respectively, and their IOP was determined using identical methods employed in the pharmacology studies. Treatment of wild-type B6.129 hybrid mice with ROCK inhibitors (Y-27632 and Y-39983) resulted in significant reductions in IOP. The magnitude of IOP reduction observed with topical Y-39983 was comparable to timolol, and exceeded the IOP effects of latanoprost in this study. Pilocarpine had no discernible effect on IOP in mice. Moreover, mice deficient in either Rock1 or Rock2 exhibited a significant decrease in IOP compared to their B6.129 wild-type littermates. Pharmacological or genetic inhibition of ROCKs results in decreased IOP in mice. The magnitude of IOP reduction is significant as demonstrated with comparative pharmacology using agents that lower IOP in humans. These studies support the ROCK pathway as a therapeutic target for treating ocular hypertension.

Research paper thumbnail of The Normal patched Allele Is Expressed in Medulloblastomas from Mice with Heterozygous Germ-Line Mutation of patched1

Cancer Research, Apr 15, 2000

Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila seg... more Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila segment polarity gene, patched (ptc) and its ligand, sonic hedgehog (shh), contribute to tumor formation in several tissues. Recently, a subset of medulloblastoma, the most common malignant brain tumor in children, was found to contain somatic mutations in the human ptc gene. In addition, basal cell nevus syndrome (BCNS), or Gorlin syndrome, which is characterized by developmental anomalies and a predisposition to skin and nervous system malignancies, is associated with germ-line mutation of ptc. Targeted disruption of both alleles of ptc in mice results in embryonic lethality. However, ptc ؉/؊ mice survive and develop spontaneous cerebellar brain tumors, suggesting that ptc may function as a tumor suppressor gene. Therefore, we investigated ptc ؉/؊ mice as a model for human medulloblastoma. We report that 14% of ptc ؉/؊ mice develop central nervous system tumors in the posterior fossa by 10 months of age, with peak tumor incidence occurring between 16 and 24 weeks of age. The tumors exhibited several characteristics of human medulloblastoma, including expression of intermediate filament proteins specific for neurons and glia. Full-length ptc mRNA was present in all tumors analyzed, indicating that there was no loss of heterozygosity at the ptc locus. Nucleotide sequence of ptc mRNA from four tumors failed to identify any mutations. However, a comparison of the normal ptc sequence from C57BL/6 and 129Sv mice did reveal several polymorphisms. High levels of gli1 mRNA and protein were detected in the tumors, suggesting that the shh/ptc pathway was activated despite the persistence of ptc expression. These data indicate that haploinsufficiency of ptc is sufficient to promote oncogenesis in the central nervous system.

Research paper thumbnail of The molecular basis of fragile X syndrome

Cold Spring Harbor symposia on quantitative biology, 1996

Research paper thumbnail of Cysteine 64 of Ref-1 is not essential for redox regulation of AP-1 DNA binding

Molecular and cellular biology, 2003

Ref-1 participates in DNA repair as well as in redox regulation of transcription factor function.... more Ref-1 participates in DNA repair as well as in redox regulation of transcription factor function. The redox function of Ref-1 involves reduction of oxidized cysteine residues within the DNA binding domains of several transcription factors, including Fos and Jun. Reduction of these residues is required for DNA binding, providing a redox-dependent mechanism for regulation of target gene expression. Previous in vitro studies implicated cysteine 65 of human Ref-1 (cysteine 64 of mouse Ref-1) as the redox catalytic site. We analyzed the in vivo role of cysteine 64 in redox regulation of AP-1 activity by introducing a cysteine-to-alanine point mutation into the endogenous mouse Ref-1 gene (ref-1(C64A)). Unlike Ref-1 null mice, which die very early in embryonic development, homozygous ref-1(C64A) mice are viable, they survive to normal life expectancy, and they display no overt abnormal phenotype. Although Ref-1 provides the major AP-1-reducing activity in murine cells, ref-1(C64A) cells r...

Research paper thumbnail of Loss of p53 but not ARF accelerates medulloblastoma in mice heterozygous for patched

Cancer research, Jan 15, 2001

Brain malignancies represent the most common solid tumors in children, and they are responsible f... more Brain malignancies represent the most common solid tumors in children, and they are responsible for significant mortality and morbidity. The molecular basis of the most common malignant pediatric brain tumor, medulloblastoma, is poorly understood. Mutations in several genes including the human homologue of the Drosophila segment polarity gene, patched (PTCH), the adenomatous polyposis coli gene (APC), beta-catenin, and p53 have been reported in subsets of hereditary and sporadic medulloblastoma. Inactivation of one Ptc allele in mice results in a 14% incidence of medulloblastoma. Here, we report a dramatic increase in the incidence (>95%) and accelerated development (prior to 12 weeks of age) of medulloblastoma in mice heterozygous for Ptc that lack p53. The acceleration of tumorigenesis in Ptc+/- mice is specific for loss of p53, because no change in tumor incidence was observed in Ptc+/- mice carrying a mutation in APC (Min+/-) or in Ptc+/- mice deficient in p19ARF. Thus, there...

Research paper thumbnail of The normal patched allele is expressed in medulloblastomas from mice with heterozygous germ-line mutation of patched

Cancer research, Jan 15, 2000

Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila seg... more Defects in a developmental signaling pathway involving mammalian homologues of the Drosophila segment polarity gene, patched (ptc) and its ligand, sonic hedgehog (shh), contribute to tumor formation in several tissues. Recently, a subset of medulloblastoma, the most common malignant brain tumor in children, was found to contain somatic mutations in the human ptc gene. In addition, basal cell nevus syndrome (BCNS), or Gorlin syndrome, which is characterized by developmental anomalies and a predisposition to skin and nervous system malignancies, is associated with germ-line mutation of ptc. Targeted disruption of both alleles of ptc in mice results in embryonic lethality. However, ptc+/- mice survive and develop spontaneous cerebellar brain tumors, suggesting that ptc may function as a tumor suppressor gene. Therefore, we investigated ptc+/-mice as a model for human medulloblastoma. We report that 14% of ptc+/- mice develop central nervous system tumors in the posterior fossa by 10 mo...

Research paper thumbnail of Fragile X mental retardation protein: nucleocytoplasmic shuttling and association with somatodendritic ribosomes

The Journal of neuroscience : the official journal of the Society for Neuroscience, 1997

Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unsta... more Fragile X syndrome, a leading cause of inherited mental retardation, is attributable to the unstable expansion of a CGG-repeat within the FMR1 gene that results in the absence of the encoded protein. The fragile X mental retardation protein (FMRP) is a ribosome-associated RNA-binding protein of uncertain function that contains nuclear localization and export signals. We show here detailed cellular localization studies using both biochemical and immunocytochemical approaches. FMRP was highly expressed in neurons but not glia throughout the rat brain, as detected by light microscopy. Although certain structures, such as hippocampus, revealed a strong signal, the regional variation in staining intensity appeared to be related to neuron size and density. In human cell lines and mouse brain, FMRP co-fractionated primarily with polysomes and rough endoplasmic reticulum. Ultrastructural studies in rat brain revealed high levels of FMRP immunoreactivity in neuronal perikarya, where it is co...