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Biomedical subjects

K Farrell

Publications and source records attributed to K Farrell.

At least 37 records · Page 2Linked to original sources
PubMed1997

Neuronal regulation of glutamate transporter subtype expression in astrocytes.

GLT-1, GLAST, and EAAC1 are high-affinity, Na(+)-dependent glutamate transporters identified in rat forebrain. The expression of these transporter subtypes was characterized in three preparations: undifferentiated rat cortical astrocyte cultures, astrocytes cocultured with cortical neurons, and astrocyte cultures differentiated with dibutyryl cyclic AMP (dBcAMP). The undifferentiated astrocyte monocultures expressed only the GLAST subtype. Astrocytes cocultured with neurons developed a stellate morphology and expressed both GLAST and GLT-1; neurons expressed only the EAAC1 transporter, and rare microglia in these cultures expressed GLT-1. Treatment of astrocyte cultures with dBcAMP induced expression of GLT-1 and increased expression of GLAST. These effects of dBcAMP on transporter expression were qualitatively similar to those resulting from coculture with neurons, but immunocytochemistry showed the pattern of transporter expression to be more complex in the coculture preparations. Compared with astrocytes expressing only GLAST, the dBcAMP-treated cultures expressing both GLAST and GLT-1 showed an increase in glutamate uptake Vmax, but no change in the glutamate K(m) and no increased sensitivity to inhibition by dihydrokainate. Pyrrolidine-2,4-dicarboxylic acid and threo-beta-hydroxyaspartic acid caused relatively less inhibition of transport in cultures expressing both GLAST and GLT-1, suggesting a weaker effect at GLT-1 than at GLAST. These studies show that astrocyte expression of glutamate transporter subtypes is influenced by neurons, and that dBcAMP can partially mimic this influence. Manipulation of transporter expression in astrocyte cultures may permit identification of factors regulating the expression and function of GLAST and GLT-1 in their native cell type.

ATP-Binding Cassette Transporters↗
PubMed1997

Astrocyte energetics, function, and death under conditions of incomplete ischemia: a mechanism of glial death in the penumbra.

Cerebral artery occlusion produces regions of incomplete ischemia (the ischemic penumbra), which, in the absence of reflow, undergo progressive metabolic deterioration culminating in infarction. The factors causing infarction are not yet established, but progression to cell death is preceded by progressive acidosis, decreasing glucose utilization, and ATP depletion. To identify potential mechanisms of glial death in the ischemic penumbra, astrocytes in culture were subjected to conditions that occur during incomplete ischemia: hypoxia, acidosis, and raised extracellular K+. Neither acidosis (to pH 6.2) nor chemical hypoxia (5 mM azide) alone produced significant astrocyte death or marked ATP depletion. By contrast, hypoxia combined with acidosis caused near-complete ATP depletion by 3.5 h and 70% cell death after 7 h. Glycolytic rate increased during hypoxia alone but decreased during hypoxia with acidosis. Since glycolysis is the sole source of ATP production during hypoxia, acidosis inhibition of glycolysis is a likely cause of the far greater ATP depletion resulting from hypoxia with acidosis. Glutamate uptake was reduced during hypoxia and further reduced during hypoxia with acidosis, consistent with the changes in astrocyte ATP. Glutamate uptake, ATP levels, and glycolytic rate each exhibited reductions that were progressive over 3 h of hypoxia with acidosis, and these changes were accompanied by progressive intracellular acidosis. Since ATP depletion leads to acidosis, and acidosis inhibits glycolysis, these findings suggest a regenerative cycle initiated by the combination of hypoxia with acidosis. This cycle could result in progressive metabolic decline and cell death in the ischemic penumbra.

Acidosis↗
PubMed1997

The S factor--a new derived hemodynamic oxygenation parameter--a useful tool for simplified mathematical modeling of global problems of oxygen transport.

We describe a new derived hemodynamic oxygenation parameter, the S factor (S). The factor is based on oxygen delivery and oxygen consumption and can range from -3 to 1. It allows simplified mathematical modeling of clinical problems of oxygen transport and can be applied to many clinical situations. A new hemodynamic oxygenation parameter, the S factor (S), is introduced as an aid to mathematical modeling. It is defined as follows: [formula: see text] (DO2 = oxygen delivery, VO2 = oxygen consumption) S can theoretically vary from -3 (DO2 = VO2) to +1 (VO2 = 0). When DO2/VO2 = 4 (ie. OER = 0.25), S = 0. An S < 0 implies utilization of reserve oxygen transport capacity. An S > 0 implies increased oxygen delivery in relation to oxygen consumption (ie. "shunted oxygen delivery"). By algebraic manipulation and substitution of the components of DO2 into Equation 1: DO2 = Q x Ca x 10 DO2 = Q [(Hb)(Sat)(1.36) + PaO2(.0031)] 10 (2) the following equations can be derived: [formula: see text] [formula: see text] Ca - Cv (Ca = arterial content, Cv = venous content) can be determined by substituting components of oxygen consumption: VO2 = Q (Ca - Cv) x 10 (5) into equation 1 and solving for Ca - Cv. [formula: see text] Equation 6 can be simplified to: [formula: see text] A previously defined relationship between mixed venous PO2 (PvO2) and DO2/VO2 (where calculated P50 is 26.6 +/- 1.0) can be used to modify S in a clinically relevant manner. PvO2 = 5.44D O2/VO2 + 18.16 (8) The relationship between S and PvO2 can be defined by substituting Equation 4 into Equation 1 and solving for PvO2 PvO2 = [21.76/(1-S)] + 18.16 (9) As an example, at a PvO2 of 28 torr (anaerobic threshold), S = -1.2. The relationship between PvO2 and S is shown in Figure 1. S, which can also be defined as 1-4(VO2/DO2) or 1-4(OER), is a useful tool for mathematical modeling of global problems of oxygen transport because the previously derived equations with the S value allow the components of oxygen transport to be interrelated in a clinically relevant manner. Additional advantages of using S in mathematical modeling are: 1. Conceptually it 'fits' in that in regards to the sign (+ or -), as a -S implies utilization of reserve oxygen transport capacity and a +S implies wasted or excess oxygen delivery (shunted). 2. These concepts are easily quantified using the S factor. 3. It 'spreads out' the difference between values for parameters (OER or S) integrating components of oxygen transport, ie. in the 'normal state' regarding oxygen transport, OER = 0.25 and S = 0. At the anaerobic threshold (PvO2 = 28 torr), OER = 0.55 and S = -1.2. Thus, the change in OER from 'normal state' to anaerobic threshold is 0.3 (0.55-0.25) and the change in S is 1.2. This represents a four-fold increase. Four examples of mathematical modeling of global problems of oxygen transport using the S factor are described below.

Anaerobiosis↗
PubMed1997

Prospective, open-label, add-on study of lamotrigine in 56 children with intractable generalized epilepsy.

The role of lamotrigine (LTG) in childhood epilepsy is emerging. We evaluated the efficacy and adverse effects of LTG in an open, prospective study of 56 children with generalized epilepsies. Six (11%) children became seizure-free, and 24 (43%) had greater than 50% reduction in seizure frequency. LTG was effective against a broad range of generalized seizure types. Three of 15 patients with Lennox-Gastaut syndrome achieved complete seizure control and eight demonstrated 50 to 99% improvement in seizure control. Increase in seizures (7) and rash (5) were the most common side effects. After valproate was discontinued, LTG therapy was resumed, with no recurrence of rash in any patient. This study suggests that LTG may be a useful drug in the treatment of generalized epilepsies in children.

Adolescent↗
PubMed1997

Unstable insertion in the 5' flanking region of the cystatin B gene is the most common mutation in progressive myoclonus epilepsy type 1, EPM1.

Progressive myoclonus epilepsy type 1 (EPM1, also known as Unverricht-Lundborg disease) is an autosomal recessive disorder characterized by progressively worsening myoclonic jerks, frequent generalized tonic-clonic seizures, and a slowly progressive decline in cognition. Recently, two mutations in the cystatin B gene (also known as stefin B, STFB) mapping to 21q22.3 have been implicated in the EPM1 phenotype: a G-->C substitution in the last nucleotide of intron 1 that was predicted to cause a splicing defect in one family, and a C-->T substitution that would change an Arg codon (CGA) to a stop codon (TGA) at amino acid position 68, resulting in a truncated cystatin B protein in two other families. A fourth family showed undetectable amounts of STFB mRNA by northern blot analysis in an affected individual. We present haplotype and mutational analyses of our collection of 20 unrelated EPM1 patients and families from different ethnic groups. We identify four different mutations, the most common of which consists of an unstable approximately 600-900 bp insertion which is resistant to PCR amplification. This insertion maps to a 12-bp polymorphic tandem repeat located in the 5' flanking region of the STFB gene, in the region of the promoter. The size of the insertion varies between different EPM1 chromosomes sharing a common haplotype and a common origin, suggesting some level of meiotic instability over the course of many generations. This dynamic mutation, which appears distinct from conventional trinucleotide repeat expansions, may arise via a novel mechanism related to the instability of tandemly repeated sequences.

Base Sequence↗
PubMed1997

Intrathecally administered baclofen for treatment of children with spasticity of cerebral origin.

Management of severe spasticity in children is often a difficult problem. Orally administered medications generally offer limited benefits. This study examines the value of intrathecally administered baclofen in the treatment of 19 children with severe spasticity of cerebral origin: eight of whom sustained brain injury associated with trauma, near drowning, or cardiac arrest; 10 with cerebral palsy (spastic quadriplegia); and one child with Leigh's disease. At the time of entry into the study, patients ranged from 4 to 19 years of age, and all were completely dependent on caretakers for activities of daily living. Children who responded positively to a trial dose of intrathecal baclofen underwent insertion of a drug delivery system for continuous infusion. This was followed by a double-blind trial of baclofen or placebo and follow-up review at 3 and 6 months, and yearly thereafter. Seven children did not undergo pump implantation because of excess sedation or poor response. The 12 remaining children have been followed for a period of 1 to 5 years. Favorable responses were present in all 12 children as determined by the Ashworth Scale, with the greatest benefit being reduction of lower limb tone. Except in the case of one child who had reduction in lower limb tone that resulted in difficulty with transfers, the caretakers all reported significant benefits from intrathecal baclofen, with improvement in muscle tone, behavior, sitting, and general ease of care being most commonly noted. Central side effects were seen in some children who received continuous intrathecal baclofen infusion and included hypotension (two patients), bradycardia (two), apnea or respiratory depression (two), and sedation (one). During a total of 568 months of pump operation there were 10 mechanical complications, including two related to pump or side port failure and eight related to catheter kinks, extrusions, or dislodgment. Pump pocket effusion occurred in five children and a cerebrospinal fluid fistula was seen in one child. Local infection occurred in three children and meningitis in two children. The results demonstrate the potential value of continuous intrathecal baclofen infusion for treatment of severe spasticity of cerebral origin. However, this treatment can result in significant complications and more experience is required before the long-term benefits can be determined.

Adolescent↗
PubMed1997

A rapid in vitro assay for evaluation of metabolism-dependent cytotoxicity of antiepileptic drugs on isolated human lymphocytes.

In vitro assessment of human lymphocyte viability by trypan blue dye exclusion in the presence of an external metabolizing system (microsomes plus NADPH) has been shown to be a useful method in assessing predisposition to idiopathic toxicity in response to various anticonvulsant drugs. The trypan blue method, however, is labor intensive, is time consuming, is prone to human error, is not suitable for high-volume toxicity screening, and excludes autolysed cells. The objective of this study was to develop a rapid, high-capacity, objective, and easy in vitro cytotoxicity method for the detection of metabolism-dependent cytotoxicity of a test chemical. The in vitro system uses an external metabolizing system (rabbit microsomes) in conjunction with isolated human lymphocytes as the target cells. Cellular toxicity was determined by assessing plasma membrane integrity using a membrane-impermeant fluorescent nucleic acid dye (YO-PRO-1) and a multiwell plate scanner for fluorescence. Using this system, cells incubated with either acetaminophen (1500 micrograms/ml), carbamazepine (62.5 microM), phenytoin (62.5 microM), or phenobarbital (62.5 microM) showed net increases in percentage cell death of 31 +/- 5, 11 +/- 4, 0 +/- 3, and 2 +/- 3, respectively. A metabolism-dependent concentration-response was observed for valproic acid-induced cytotoxicity, which approached a plateau at a concentration of 4000 micrograms/ml with a net percentage cell death of 31 +/- 4. This technique resolves various technical difficulties inherent in viability determinations by the trypan blue exclusion method. The YO-PRO-1 method also may be useful in a clinical setting for the assessment of patients with a genetically determined susceptibility to certain drugs and for identifying the responsible drug in patients with idiopathic toxicity undergoing multiple-drug therapy.

Acetaminophen↗
PubMed1995

Acidosis causes failure of astrocyte glutamate uptake during hypoxia.

Failure of glutamate uptake during ischemia can lead to neurotoxic accumulations of glutamate in brain extracellular space. Hypoxia and acidosis are metabolic consequences of ischemia that may individually or in combination impair glutamate uptake. We used primary rat astrocyte cultures to study the effects of acidosis, chemical hypoxia, and the combination of acidosis plus chemical hypoxia on glutamate uptake. Chemical hypoxia alone reduced uptake by 35-45%. Reduction in pH from 7.4 to 5.8 also caused a significant but incomplete inhibition of glutamate uptake, and this effect was more pronounced in medium buffered with CO2/bicarbonate. However, the combination of chemical hypoxia plus acidosis reduced glutamate uptake to below 10% of controls. Astrocyte ATP levels, like glutamate uptake, were significantly reduced by chemical hypoxia and further reduced by the combination of hypoxia plus acidosis. Acidosis under normoxic conditions had no significant effect on astrocyte ATP levels. These results suggest two mechanisms by which acidosis may contribute to failure of astrocyte glutamate uptake during ischemia: Acidosis may act in concert with hypoxia to cause ATP depletion, and acidosis may also have direct effects on glutamate transporters unrelated to effects on cellular ATP levels. pH effects on glutamate uptake may be an important factor affecting neuronal survival during incomplete ischemia.

Acidosis↗
PubMed1995

Seizures involving the supplementary sensorimotor area in children: a video-EEG analysis.

Most reports of supplementary sensorimotor seizures have been of adults with medically refractory epilepsy. Typically, supplementary sensorimotor seizures have onset in childhood. We describe the electroclinical features in 12 children. Cases were selected from an EEG laboratory population in whom video-EEG was performed routinely on all children. Supplementary sensorimotor seizures were diagnosed when there was bilateral tonic posturing of the upper or lower extremities, preserved consciousness, and lack of postictal confusion. Sensory auras were reported by 8 of 9 children aged > 3 years who had daytime seizures. Speech arrest occurred in all patients in whom it could be assessed, and abnormal vocalization was observed in 7 children. Interictal EEGs were often normal (49% of recordings), and ictal EEG changes could be subtle. An etiology was demonstrated in only 2 children, and brain imaging studies were normal in the other 10 patients. Seizures were controlled with antiepileptic drugs (AEDs) in 6 of the 12 children. The clinical manifestations of supplementary sensorimotor seizures in children are similar to those reported in adults; misdiagnosis is common at all ages.

Age Factors↗
PubMed1994

Genetic control of programmed cell death in Drosophila.

A gene, reaper (rpr), that appears to play a central control function for the initiation of programmed cell death (apoptosis) in Drosophila was identified. Virtually all programmed cell death that normally occurs during Drosophila embryogenesis was blocked in embryos homozygous for a small deletion that includes the reaper gene. Mutant embryos contained many extra cells and failed to hatch, but many other aspects of development appeared quite normal. Deletions that include reaper also protected embryos from apoptosis caused by x-irradiation and developmental defects. However, high doses of x-rays induced some apoptosis in mutant embryos, and the resulting corpses were phagocytosed by macrophages. These data suggest that the basic cell death program is intact although it was not activated in mutant embryos. The DNA encompassed by the deletion was cloned and the reaper gene was identified on the basis of the ability of cloned DNA to restore apoptosis to cell death defective embryos in germ line transformation experiments. The reaper gene appears to encode a small peptide that shows no homology to known proteins, and reaper messenger RNA is expressed in cells destined to undergo apoptosis.

Amino Acid Sequence↗
PubMed1994

Cranial neuropathy, polyneuropathy and thrombocytopenia with Epstein-Barr virus infection.

Neurological involvement is an uncommon complication of Epstein-Barr virus infection, and the long incubation period may complicate the diagnosis. A 15-year-old boy is described with Epstein-Barr virus infection complicated by prolonged life-threatening thrombocytopenia, cranial neuropathy and peripheral sensorimotor neuropathy. The abnormal platelet count was unresponsive to multiple-drug therapy and splenectomy, but normalized 13 months after presentation. Neurological recovery was slow; the patient continues to have a mild peripheral sensorimotor neuropathy three years after the onset of the illness. Infection with Epstein-Barr virus should be considered in patients with acute neurological problems associated with thrombocytopenia. Measurement of antibodies to individual Epstein-Barr virus proteins facilitates the diagnosis of Epstein-Barr virus infection.

Adolescent↗
PubMed1994

Glycosylation-dependent inactivation of the ecotropic murine leukemia virus receptor.

The ecotropic murine leukemia virus (E-MuLV) receptor expressed on Mus dunni tail fibroblast (MDTF) cells is a receptor for all E-MuLVs with the notable of Moloney murine leukemia virus (Mo-MuLV). Substitution of isoleucine for valine at position 214 in the third extracellular region (the putative E-MuLV binding site) of the MDTF receptor molecule allows this molecule to function as a Mo-MuLV receptor (M.V. Eiden, K. Farrell, J. Warsowe, L. A. Mahan, and C. A. Wilson, J. Virol. 67:4056-4061, 1993). We have now determined that treating MDTF cells with tunicamycin, an inhibitor of N-linked glycosylation, also renders them susceptible to Mo-MuLV infection. Two potential N-linked glycosylation sites are present in the third extracellular regions of both the NIH 3T3 and MDTF ecotropic receptors. The glycosylation site at position 229 of the MDTF receptor cDNA was eliminated by substituting a threonine codon for the asparagine codon. Mo-MuLV-resistant human HOS cells, expressing this form of the receptor, are susceptible to Mo-MuLV infection. Thus, our studies suggest that without a glycan moiety at position 229, the valine residue at 214 is no longer restrictive for Mo-MuLV infection. BHK-21 and CHO K1 hamster cells also express glycosylation-inactivated forms of the ecotropic receptor. Sequence analysis of these receptors together with our analysis of MDTF receptor function suggests that a single asparagine-linked glycosylation site is responsible for glycosylation inactivation of these receptors.

Amino Acid Sequence↗
PubMed1994

Sweat gland vacuoles in Unverricht-Lundborg disease: a clue to diagnosis?

There has been no reported pathologic abnormality outside the central nervous system in patients with Unverricht-Lundborg disease (ULD). We report membrane-bound vacuoles with clear contents in eccrine clear cells and dark cells in five of seven patients with ULD, as well as in one clinically unaffected sibling. Vacuoles were not seen in the biopsies of two patients and of eight controls with Lafora's disease. These findings, though not entirely specific, suggest that skin biopsy may serve as a diagnostic aid to give supportive evidence for ULD.

Adolescent↗