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R A Fenstermaker

Publications and source records attributed to R A Fenstermaker.

33 records · Page 2Linked to original sources
PubMed1993

Transcriptional control of the platelet-derived growth factor subunit genes.

Platelet-derived growth factor (PDGF) is a potent mitogen consisting of heterodimers of two distinct but homologous polypeptide chains, denoted A and B. PDGF-like homodimers of the A- and B-chains have been isolated, as well as two distinct receptor types (alpha and beta), which discriminate among the PDGF isoforms. The PDGF A- and B-chains are encoded by distinct genes located on human chromosomes 7 and 22, respectively. Although PDGF has been implicated as an important participant in development, tissue repair, and numerous pathologic states including tumorigenesis, atherosclerosis and inflammation, the mechanisms which determine the rate of its synthesis are only beginning to be understood. Basal expression of the PDGF A- and B-chain genes has been characterized in a number of cell types and is directed in part by elements in the respective proximal promoter-regulatory regions of the two genes. In addition, the first intron of PDGF-B has been shown to contain both positive and negative regulatory elements. Transcription of the PDGF subunit genes is inducible by a wide variety of mitogenic growth factors, cytokines and other agonists. These agents produce a rapid increase in steady-state concentrations of PDGF A- and B-chain mRNAs, peaking within 4-8 h of stimulation. The inductive effects of protein kinase C-activating phorbol 12-myristate 13-acetate (PMA), thrombin and transforming growth factor-beta (TGF-beta) are mediated through increases in the transcription rates of both genes. In addition, cAMP blocks the increases in transcription of the B-chain gene induced by thrombin and TGF-beta. Studies have demonstrated the importance of sequences immediately upstream of the B-chain transcription start site for induction in response to PMA-initiated megakaryocyte differentiation, an effect which is dependent on protein synthesis. However, cis-acting elements which mediate more rapid transcriptional induction seen in endothelial cells and astrocytes have yet to be identified in the proximal 5'-flanking sequences of either the A- or B-chain genes, suggesting that such events may be mediated by elements located outside of this region.

Base Sequence↗
PubMed1993

Acute neurologic management of the patient with spinal cord injury.

Early and effective spinal immobilization, physiological support, precise radiographic analysis, and surgical intervention tailored to the specific anatomic pathology form the basis for appropriate management of spinal injury. Despite improvements in each of these areas, however, the neurologic outcome in patients with severe deficits remains problematic. Research into the prevention of adverse metabolic changes that occur in the acutely injured spinal cord may offer hope for improved outcomes.

Acute Disease↗
PubMed1991

Different combinations of regulatory elements may explain why placenta-specific expression of the glycoprotein hormone alpha-subunit gene occurs only in primates and horses.

Expression of the glycoprotein hormone alpha-subunit gene occurs in the pituitary of all mammals but in placenta of only primates and horses. In humans, two different elements, termed upstream regulatory element (URE) and cAMP response element (CRE), are required for placenta-specific expression of the alpha-subunit gene. The URE binds a protein unique to placenta whereas the CRE binds a ubiquitous protein. Comparative analysis of the promoter-regulatory region of the alpha-subunit gene from a number of mammals indicates that a functional URE has been retained and suggests the potential for placenta-specific expression. Indirect evidence also indicates that the URE-binding protein has been conserved, even in placenta from mammals that fail to express the alpha-subunit gene. Lack of expression of the alpha-subunit gene in placenta of rodents and cattle can be traced to a single nucleotide change that renders the CRE-like sequence of these genes incapable of binding the protein that confers responsiveness to cAMP. In contrast, although expression of the alpha-subunit gene occurs in horse placenta, the promoter-regulatory region lacks a functional CRE but appears to retain a functional URE. This suggests that either a different accessory element and cognate protein interacts with the horse URE to provide placenta-specific expression or that a completely different set of regulatory elements is required for placenta-specific expression in horses.

Animals↗
PubMed1990

Different combinations of regulatory elements may account for expression of the glycoprotein hormone alpha-subunit gene in primate and horse placenta.

Expression of the glycoprotein hormone alpha-subunit gene occurs in the pituitaries of all mammals and in the placentas of primates and horses. In humans, tandem cAMP response elements (CREs), located in the proximal promoter-regulatory region of the alpha-subunit gene, act together with an adjacent upstream regulatory element to confer placenta-specific expression. Here, we report that the alpha-subunit genes of Old World Monkeys contain a single functional CRE. This suggests that tandem CREs are unique to higher primates and humans and are not absolutely required for placenta-specific expression. In contrast, the comparable promoter-regulatory region of the horse alpha-subunit gene lacks a functional CRE but appears to retain a functional upstream regulatory element. This suggests that acquisition of placenta-specific expression of the alpha-subunit gene occurred independently in these distantly related mammals. As a result, different combinations of cis-acting elements may explain why expression of the alpha-subunit gene only occurs in placenta of primates and horses.

Animals↗
PubMed1989

CRE-binding proteins interact cooperatively to enhance placental-specific expression of the glycoprotein hormone alpha-subunit gene.

The 18-bp direct repeat occurring between positions -146 and -111 in the 5'-flanking region of the human alpha-subunit gene serves two functions: it mediates the transcriptional effect of cAMP and it acts in conjunction with an adjacent cis-acting element (URE) to confer properties of placental-specific expression to the alpha-subunit promoter. Functional activity of the URE and CRE requires binding of a trans-acting factor; each element binds a different factor. Analysis of saturation isotherms provides good evidence that cooperativity is involved in binding of CREB to the 18-bp direct repeat. This cooperativity could account for the synergistic effect of two CRE on both basal and cAMP-stimulated transcription. It remains to be determined whether heterotropic cooperativity is involved in binding of trans-acting factors to the URE and CRE. A major difference between the 5'-flanking region of the human alpha-subunit gene and comparable regions from bovine, rat, and mouse alpha-subunit genes is that the latter contain a single CRE homolog which appears incapable of binding the trans-acting factor that binds to the human alpha CRE. Lack of a functional CRE provides at least one explanation for inactivity of the bovine alpha-subunit promoter in choriocarcinoma cells and probably in bovine placenta as well. Yet, the same bovine promoter-regulatory region that lacks a functional CRE is capable of conferring pituitary-specific expression to the CAT gene in transgenic mice (data not shown). This suggests that the CRE is not required for pituitary-specific expression of the bovine alpha-subunit gene. Instead, another cis-acting element(s) must confer this property to the alpha-subunit promoter. While it is tempting to suggest that bovine, rat, and mouse alpha-subunit genes are not regulated by cAMP because of their inactive CRE homolog, it is also quite possible that other CRE are located further upstream. Accordingly, it will be of interest to obtain additional 5'-flanking sequence and determine whether functional homologs of the human alpha CRE are present in the bovine, rat, and mouse alpha-subunit genes, or whether another class of cis-acting elements provide cAMP-responsiveness.

Base Sequence↗
PubMed1989

Expression of the glycoprotein hormone alpha-subunit gene in the placenta requires a functional cyclic AMP response element, whereas a different cis-acting element mediates pituitary-specific expression.

The single-copy gene encoding the alpha subunit of glycoprotein hormones is expressed in the pituitaries of all mammals and in the placentas of only primates and horses. We have systematically analyzed the promoter-regulatory elements of the human and bovine alpha-subunit genes to elucidate the molecular mechanisms underlying their divergent patterns of tissue-specific expression. This analysis entailed the use of transient expression assays in a chorionic gonadotropin-secreting human choriocarcinoma cell line, protein-DNA binding assays, and expression of chimeric forms of human or bovine alpha subunit genes in transgenic mice. From the results, we conclude that placental expression of the human alpha-subunit gene requires a functional cyclic AMP response element (CRE) that is present as a tandem repeat in the promoter-regulatory region. In contrast, the promoter-regulatory region of the bovine alpha-subunit gene, as well as of the rat and mouse genes, was found to contain a single CRE homolog that differed from its human counterpart by a single nucleotide. This difference substantially reduced the binding affinity of the bovine CRE homolog for the nuclear protein that bound to the human alpha CRE and thereby rendered the bovine alpha-subunit promoter inactive in human choriocarcinoma cells. However, conversion of the bovine alpha CRE homolog to an authentic alpha CRE restored activity to the bovine alpha-subunit promoter in choriocarcinoma cells. Similarly, a human but not a bovine alpha transgene was expressed in placenta in transgenic mice. Thus, placenta-specific expression of the human alpha-subunit gene may be the consequence of the recent evolution of a functional CRE. Expression of the human alpha transgene in mouse placenta further suggests that evolution of placenta-specific trans-acting factors preceded the appearance of this element. Finally, in contrast to their divergent patterns of placental expression, both the human and bovine alpha-subunit transgenes were expressed in mouse pituitary, indicating differences in the composition of the enhancers required for pituitary- and placenta-specific expression.

Animals↗
PubMed1989

The transcriptional response of the human chorionic gonadotropin beta-subunit gene to cAMP is cycloheximide sensitive and is mediated by cis-acting sequences different from that found in the alpha-subunit gene.

Cyclic AMP stimulates transcription of the genes encoding the alpha- and beta-subunits of human CG (hCG). Although the cis-acting cAMP response element (CRE) of the alpha-subunit gene has been extensively characterized, relatively little is known about the putative response element of the hCG beta gene. Here, we demonstrate that cAMP stimulation of hCG beta gene transcription requires ongoing protein synthesis, whereas cAMP stimulation of alpha-subunit gene transcription does not. These results suggest that cAMP-stimulated transcription of the alpha and hCG beta genes may involve different cis-acting elements and trans-acting factors. Accordingly, we have constructed a series of deletion mutants consisting of fragments of the hCG beta promoter-regulatory region linked to the bacterial chloramphenicol acetyl transferase gene. To potentiate detection of a CRE, we constructed vectors containing the Rous sarcoma virus enhancer to augment transcriptional activity of the hCG beta-promoter. We also used vectors designed to determine whether regions containing a putative CG beta CRE could confer cAMP responsiveness to a heterologous promoter. These constructs were evaluated for activity by transfection into choriocarcinoma (BeWo) cells. Transient expression of these vectors identifies a broad region which renders transcription of the chloramphenicol acetyl transferase gene responsive to cAMP. Surprisingly, division of the hCG beta 5'-flanking and untranslated regions into smaller fragments led to loss of cAMP responsiveness, suggesting that the hCG beta CRE may be comprised of multiple domains dispersed across the proximal 650 base pairs of CG beta 5'-flanking and untranslated sequence.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Preservation of hearing in the surgical removal of cerebellopontine angle tumors.

It is claimed that postsurgical hearing preservation is possible in about 5 to 8 per cent of cases of acoustic neuroma. Even with small tumors, hearing preservation can be accomplished in about half of the patients who are fortunate to have anatomic integrity of facial and cochlear nerves, as well as intact inner ear blood supply at the end of the surgical procedure. Monitoring of seventh and eighth nerve function through evoked potentials may be important. On the other hand, even if evoked potentials are preserved during surgery with wave V latency and amplitude similar to preoperative recording, hearing may still be completely lost during the immediate postoperative period. There is no predictable pattern that assures postoperative preservation of hearing. In Case 2, the seventh and eighth nerves as well as the inner ear blood supply were carefully dissected and anatomically and electrophysiologically preserved during the intraoperative period. At the end of the surgical procedure, the evoked potentials were the same as preoperatively. Nevertheless, the patient ended up with no detectable hearing postoperatively. We feel that the suboccipital-retrosigmoid transcanal approach can be safely used for the removal of cerebellopontine angle tumors of all sizes. We believe the argument that "only the translabyrinthine approach can accomplish total tumor removal" is not valid. No complications have been attributed by having the patient in the semisitting position. Older patients, who cannot tolerate the semisitting position, are operated on while in the park bench position. Planned subtotal removal of a CPA tumor is done with the patient in the supine position (transmastoid-retrolabyrinthine or retrosigmoid). The relationship between the otologic surgeon and neurosurgeon is very important. The surgical approach used should be selected on the basis of the combined experience of the surgical team. Finally, an attempt should be made to preserve facial nerve function and hearing in all suitable patients. The postoperative course using the suboccipital-retrosigmoid approach is usually benign, and the patients are discharged from the hospital between 7 and 10 days following surgery. The translabyrinthine approach, in our opinion, should be reserved for smaller lesions in patients with anacusis or with residual hearing that is not worth saving.

Adult↗
PubMed1984

Fourth ventriculoceles with extracranial extension.

The radiographic features and long-term clinical outcome in three patients who presented at birth with a cystic suboccipital mass in direct communication with the fourth ventricle are reviewed. The pathological findings in a fourth infant who died are also discussed. All surviving infants were treated with cyst excision and diversion of cerebrospinal fluid. The prognosis in these children, followed from 6 to 20 years, surpasses that of the more common occipital encephalocele, for which this entity could be mistaken. The morphogenetic implications relative to more common congenital lesions in this location are discussed.

Cerebral Ventricles↗
PubMed1983

Actions of guanidine hydrochloride on cat spinal monosynaptic reflex transmission.

The effects of a single intravenous dose of guanidine hydrochloride were examined on the function of the lumbar dorsal-ventral root and triceps surae nerve-ventral root monosynaptic reflex (MSR) in unanesthetized acute spinal (C-1 sectioned) cats. Guanidine, in a dose of 25 mg/kg, consistently depressed the amplitude of the monosynaptic reflex response while a dose of 200 mg/kg always exerted a facilitatory action. Intermediate doses of 50 or 100 mg/kg produced variable actions on the amplitude of the monosynaptic reflex from one preparation to another. The differential effects of guanidine on the evoked monosynaptic reflex are postulated to represent the ability of the drug to enhance both excitatory and inhibitory transmission, the latter predominating at smaller doses and the former becoming apparent with an increasing dose of guanidine. The facilitatory and the depressant effects of guanidine on the function of spinal reflexes were slow to develop, suggesting an intracellular site of action. These results may have significance for an understanding of the possible efficacy of guanidine in motor neuron disease in man.

Animals↗
PubMedDate not supplied

Neurosarcoidosis presenting as a large suprasellar mass. Magnetic resonance imaging findings.

We present unusual magnetic resonance imaging (MRI) findings in a case of neurosarcoidosis. MRI revealed a large solitary suprasellar mass which resembled a neoplasm. The lesion was isointense and hyperintense on T1-weighted images, hypointense on T2-weighted images, and intensely homogeneously enhancing. Biopsy revealed a polymorphous inflammatory lesion with giant cells, which extended from the hypothalamus, consistent with neurosarcoidosis. The diagnosis of neurosarcoidosis should be considered in patients presenting with large midline tumor-like suprasellar mass lesions.

Adult↗
PubMedDate not supplied

Boronated epidermal growth factor as a potential targeting agent for boron neutron capture therapy of brain tumors.

In order for boron neutron capture therapy (BNCT) to be successful, a large number (approximately 10(9)) of 10B atoms must be delivered to each cancer cell in order to sustain a lethal 10B(n, alpha)7Li reaction. The majority of high grade gliomas express an amplified epidermal growth factor receptor (EGFR) gene, and increased numbers of EGFR are found on the cell surface. If a sufficiently large number of 10B atoms could be attached to EGF, the resulting bioconjugates might be useful for targeting brain tumors. In order to accomplish this, we have boronated a fourth-generation starburst dendrimer (SD) using an isocyanato polyhedral borane, Na(CH3)3NB10H8NCO. For conjugation, reactive thiol groups were introduced into the boronated SD using N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), and EGF was derivatized with m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (sMBS). Subsequent reaction of thiol groups of derivatized BSD with maleimide groups of derivatized EGF produced stable BSD-EGF bioconjugates containing approximately 960 atoms of boron per molecule of EGF. As determined by electron spectroscopic imaging, the BSD-EGF initially was bound to the cell surface membrane and then was endocytosed, which resulted in accumulation of boron in lysosomes. The favorable in vitro properties of these bioconjugates suggest that they may be useful for the in vivo targeting of EGFR positive brain tumors.

Amino Acid Sequence↗
PubMedDate not supplied

Radiolabeling of epidermal growth factor with 99mTc and in vivo localization following intracerebral injection into normal and glioma-bearing rats.

High grade gliomas may have amplified expression of the epidermal growth factor receptor (EGFR) gene c-erb-B, which often is associated with increased expression of transmembrane EGFR. The purpose of the present study was to develop a method for labeling EGF with 99mTc and to determine whether the resulting radioligand would localize, following intracerebral injection, in rats bearing EGFR-positive gliomas. EGF has a relatively low molecular mass (approximately 6 kDa) compared to monoclonal antibodies, and this has allowed smaller bioconjugates, which should diffuse more rapidly within the brain and more effectively target disseminated glioma cells, to be constructed. In the present study, EGF has been labeled with either 131I or 99mTc, and in vitro uptake of the resulting radioligand has been investigated using C6EGFR rat glioma cells, which had been transfected with the EGFR gene. Cellular uptake of 131I radioactivity peaked after approximately 30 min of incubation with [131I]EGF, following which time it declined, while 99mTc radioactivity continued to increase over a 6 h incubation with [99mTc]-EGF. To determine if radiolabeled EGF had in vivo tumor-localizing properties, C6EGFR glioma cells were implanted stereotactically into the brains of Fischer rats. Four weeks later, either 99mTc- or 131I-labeled EGF was injected intracerebrally into normal or glioma-bearing animals using the same stereotactic coordinates. External gamma scintigraphy revealed that 131I radioactivity disappeared rapidly from the brain regions of tumor-bearing animals compared to 99mTc, approximately 50% of which remained in the tumor for up to 12 h. In contrast, only approximately 20% remained in the brains of non-tumor-bearing animals after 6 h. These studies are the first to describe a method for radiolabeling EGF with 99mTc and to detect it by external scintigraphy in the brains of tumor-bearing animals.

Animals↗
PubMedDate not supplied

Focal enhancement of cranial nerve V after radiosurgery with the Leksell gamma knife: experience in 15 patients with medically refractory trigeminal neuralgia.

BACKGROUND AND PURPOSE: Gamma knife radiosurgery is an alternative for the treatment of medically refractory trigeminal neuralgia. Few reports of posttreatment MR imaging appearance of cranial nerve V exist. Our purpose was to define MR imaging characteristics in cranial nerve V after gamma knife radiosurgery. METHODS: We retrospectively reviewed MR images of 15 patients who underwent gamma knife radiosurgery for trigeminal neuralgia. Radiation doses were 35-45 Gy at the 50% isodose line. Thin-section T2-weighted images and contrast-enhanced and nonenhanced T1-weighted images were obtained the day of radiosurgery and within the next 5 mo. Images were scored for enhancement and hyperintensity on T2-weighted images. Time to follow-up imaging and radiation dose were recorded. RESULTS: Mean time to follow-up imaging was 61 +/- 29 d. Posttreatment T2-weighted images showed stable signal intensity in all cases, with radiosurgical target site enhancement in 10. All five patients whose images did not show treatment-related enhancement received radiation doses of 35 Gy. The data suggested a correlation between enhancement with radiation dose (P =.06). No correlation of enhancement with treatment response or time to follow-up existed (P >.05). CONCLUSION: The trigeminal nerve often enhances at the target site after radiosurgery. Lack of trigeminal nerve enhancement occurred only with lower doses (35 Gy at 50%). MR imaging may be useful to confirm the presence and location of the treatment site after gamma knife radiosurgery for trigeminal neuralgia.

Adult↗