Biomedical subjects

N Akhtar

Publications and source records attributed to N Akhtar.

At least 37 records · Page 2Linked to original sources

PubMed2001

Functional analysis of the yeast Ran exchange factor Prp20p: in vivo evidence for the RanGTP gradient model.

Numerous cellular processes rely on the movement of macromolecules into and out of the nucleus. The primary regulator of this movement is the small GTPase Ran. Like other small GTPases, the nucleotide-bound state of Ran is regulated by effectors that enhance the rate of nucleotide exchange or hydrolysis. Current models for vectorial nuclear transport suggest that it is the strict compartmentalization of these Ran effector molecules that generates a gradient of RanGTP between the nucleus and the cytoplasm to impart directionality to the transport process. Here we investigate the mechanism by which the Ran exchange factor is targeted to the nucleus, and test the impact of disrupting this nuclear compartmentalization on nucleocytoplasmic transport in vivo. Our results indicate that in Saccharomycces cerevisiae the nucleotide exchange factor Prp20p can be targeted to the nucleus via a classical nuclear localization sequence. This transport mechanism is dependent both on Ran and the receptor that recognizes the nuclear localization sequence, importin alpha. Mutations in the evolutionarily conserved nuclear localization sequence only partially inhibit nuclear import of Prp20p, suggesting the existence of a secondary mechanism for this critical nuclear targeting. In an in vivo test of the RanGTP gradient model, we demonstrate that overexpression of a functional cytoplasmic exchange factor inhibits cell growth and blocks both protein import and RNA export in wild-type cells that contain the endogenous nuclear Prp20 protein. Taken together, our results provide in vivo evidence for the idea that the compartmentalization of the exchange factor serves as a mechanism for establishing directional nuclear transport.

DNA-Binding Proteins↗

PubMed2001

Efficacy of the ketogenic diet in focal versus generalized seizures.

Most reports of the ketogenic diet have focused on its efficacy for generalized seizures. Few data are available regarding its effect on focal seizures. We retrospectively studied patients (mean = 7.5 years of age) with medically intractable epilepsy treated by the ketogenic diet. The predominant seizure types in each patient were classified as generalized (100 patients) or focal (34 patients) based on ictal electroencephalograms (EEGs) or seizure semiology and interictal EEG. A seizure reduction of more than 50% compared with baseline was seen in nine patients (27%) with focal seizures and 46 patients (46%) with generalized seizures at 3 months, in 10 patients (30%) with focal seizures and 46 patients (46%) with generalized seizures at 6 months, and in eight patients (24%) with focal seizures and 42 patients (42%) with generalized seizures at 12 months. Differences were not significant. Outcome tended to be better in patients younger than 12 years of age compared with the older age group, but the difference was significant at 6 months only. Our results suggest that some patients with intractable focal epilepsy may respond favorably to the ketogenic diet and that this option should be considered if epilepsy surgery is not possible.

Adolescent↗

PubMed2001

RAC1 regulates adherens junctions through endocytosis of E-cadherin.

The establishment of cadherin-dependent cell-cell contacts in human epidermal keratinocytes are known to be regulated by the Rac1 small GTP-binding protein, although the mechanisms by which Rac1 participates in the assembly or disruption of cell-cell adhesion are not well understood. In this study we utilized green fluorescent protein (GFP)-tagged Rac1 expression vectors to examine the subcellular distribution of Rac1 and its effects on E-cadherin-mediated cell-cell adhesion. Microinjection of keratinocytes with constitutively active Rac1 resulted in cell spreading and disruption of cell-cell contacts. The ability of Rac1 to disrupt cell-cell adhesion was dependent on colony size, with large established colonies being resistant to the effects of active Rac1. Disruption of cell-cell contacts in small preconfluent colonies was achieved through the selective recruitment of E-cadherin-catenin complexes to the perimeter of multiple large intracellular vesicles, which were bounded by GFP-tagged L61Rac1. Similar vesicles were observed in noninjected keratinocytes when cell-cell adhesion was disrupted by removal of extracellular calcium or with the use of an E-cadherin blocking antibody. Moreover, formation of these structures in noninjected keratinocytes was dependent on endogenous Rac1 activity. Expression of GFP-tagged effector mutants of Rac1 in keratinocytes demonstrated that reorganization of the actin cytoskeleton was important for vesicle formation. Characterization of these Rac1-induced vesicles revealed that they were endosomal in nature and tightly colocalized with the transferrin receptor, a marker for recycling endosomes. Expression of GFP-L61Rac1 inhibited uptake of transferrin-biotin, suggesting that the endocytosis of E-cadherin was a clathrin-independent mechanism. This was supported by the observation that caveolin, but not clathrin, localized around these structures. Furthermore, an inhibitory form of dynamin, known to inhibit internalization of caveolae, inhibited formation of cadherin vesicles. Our data suggest that Rac1 regulates adherens junctions via clathrin independent endocytosis of E-cadherin.

3T3 Cells↗

PubMed2001

Depression after stroke-analysis of 297 stroke patients.

Stroke is commonly followed by some form of psychiatric disturbance. Association and severity of depression at the 3rd and 12th months after stroke in patients and controls were compared. Two registers of stroke patients were maintained over 2 years period in two different centers--one in Barisal Town and other in Mitford Hospital, Dhaka. A total of 297 first-ever strokes were registered. These patients were called back at the 3rd and 12th month interview and assessment. Beck's Depression Inventory (BDI), with 10 as the cut off point for depression, was applied to 161 of 212 survivors at the 3rd month and to 156 of 195 survivors at the 12th month. At the 3rd month, 41% of patients and 18% of controls were depressed (P<.001) and the difference was maintained at the 12th month 42% versus 19% (P<.05). Univariate risk factors for depression at the 3rd month were female sex, severe prognostic score at the onset of stroke, age more than 70 years, family history of stroke and living alone after stroke. Depression was common among stroke survivors at the 3rd months and its rate did not decrease at 1 year follow-up. Depression could be decreased if post-stroke rehabilitation services are improved. After the development of depression earlier diagnosis and management help a lot to the patients and care givers.

Aged↗

PubMed2000

SGD1 encodes an essential nuclear protein of Saccharomyces cerevisiae that affects expression of the GPD1 gene for glycerol 3-phosphate dehydrogenase.

We here report the identification of the previously uncharacterized SGD1 gene, encoding a 102.8-kDa protein containing a leucine zipper region and a bipartite nuclear localization signal. Deletion of SGD1 results in loss of cell viability, while an increased dosage of SGD1 partially suppresses the osmosensitivity of pbs2 delta and hog1 delta mutants that are defective in the osmosignaling high osmolarity glycerol (HOG) mitogen-activated protein kinase pathway. The rescued mutants display a partially re-established transcriptional control of the osmostress-induced expression of GPD1, a target gene of the HOG pathway encoding NAD(+)-dependent glycerol 3-phosphate dehydrogenase, and a partially recovered hyperosmolarity-induced production of glycerol. Consistent with Sgd1p affecting the transcriptional control of GPD1, a functional green fluorescent protein tagged Sgd1p is localized to the cell nucleus.

Cell Division↗

PubMed2000

Visualizing muscle cell migration in situ.

BACKGROUND: Cell migration has been studied extensively by manipulating and observing cells bathed in putative chemotactic or chemokinetic agents on planar substrates. This environment differs from that in vivo and, consequently, the cells can behave abnormally. Embryo slices provide an optically accessible system for studying cellular navigation pathways during development. We extended this system to observe the migration of muscle precursors from the somite into the forelimb, their cellular morphology, and the localization of green fluorescent protein (GFP)-tagged adhesion-related molecules under normal and perturbed conditions. RESULTS: Muscle precursors initiated migration synchronously and migrated in broad, rather than highly defined, regions. Bursts of directed migration were followed by periods of meandering or extension and retraction of cell protrusions. Although paxillin did not localize to discernible intracellular structures, we found that alpha-actinin localized to linear, punctate structures, and the alpha5 integrin to some focal complexes and/or vesicle-like concentrations. Alterations in the expression of adhesion molecules inhibited migration. The muscle precursors migrating in situ formed unusually large, long-lived protrusions that were polarized in the direction of migration. Unlike wild-type Rac, a constitutively active Rac localized continuously around the cell surface and promoted random protrusive activity and migration. CONCLUSIONS: The observation of cellular migration and the dynamics of molecular organization at high temporal and spatial resolution in situ is feasible. Migration from the somite to the wing bud is discontinuous and not highly stereotyped. In situ, local activation of Rac appears to produce large protrusions, which in turn, leads to directed migration. Adhesion can also regulate migration.

Animals↗

PubMed2000

Cell vacuolation induced by the VacA cytotoxin of Helicobacter pylori is regulated by the Rac1 GTPase.

Chronic gastric infection with the Gram-negative bacterium Helicobacter pylori is a major contributing factor in the development of duodenal ulcers and is believed to be a significant risk factor in the development of gastric tumors. The VacA cytotoxin of H. pylori is a 90-kDa secreted protein that forms trans-membrane ion channels. In epithelial cells, VacA activity is associated with the rapid formation of acidic vacuoles enriched for late endosomal and lysosomal markers. Rac1 is a member of the Rho family of small GTP-binding proteins that regulate reorganization of the actin cytoskeleton and intracellular signal transduction and are being shown increasingly to play a role in membrane trafficking events. In this study we report that: (i) green fluorescent-tagged Rac1 localizes around the perimeter of the vacuoles induced by VacA; (ii) expression of dominant negative Rac1 in epithelial cells inhibits vacuole formation; (iii) expression of constitutively active Rac1 potentiates the activity of VacA. Taken together, these data demonstrate a role for Rac1 in the regulation of VacA activity.

Animals↗

PubMed2000

Angiogenesis assays: problems and pitfalls.

Many studies of angiogenesis inducers and inhibitors rely on in vitro or in vivo models as indicators of efficacy. However, as valuable as these models are, there are limitations to each one of these. This overview describes some of the principal methods now in use and discusses the advantages and some of the limitations of these methods. It is suggested that multiple assays, involving both in vitro and in vivo assays, are at present the best way to minimize the problems inherent in any specific assay.

Angiogenesis Inhibitors↗

PubMed2000

Co-localization of Rac1 and E-cadherin in human epidermal keratinocytes.

The Rac1 small GTP-binding protein is known to be involved in reorganization of the actin cytoskeleton and in regulation of intracellular signal transduction. The assembly and maintenance of cadherin-based cell cell junctions in epidermal keratinocytes is thought to be dependent on activity of Rac1. In this study we have generated green fluorescent protein (GFP)-tagged wild type, dominant negative and constitutively active Rac1 expression vectors and analyzed distribution of Rac1 following microinjection of human SCC12F epidermal keratinocytes. Wild type, dominant negative and constitutively active GFP Rac1 proteins distribute to sites of cell cell adhesion and co-localize with E-cadherin and the catenins. Disruption of cadherin-based junctions by reduction in extracellular calcium concentrations, or by use of antibodies to E-cadherin, results in redistribution of Rac1 away from sites of cell cell interaction but the co-localization with E-cadherin is maintained. In addition, expression of constitutively active GFP Rac1 results in formation of membrane ruffles on the apical surface of cells and intracellular vesicles. Interestingly, co-localization of Rac1 with E-cadherin is maintained in these structures. In contrast to previously published work we find that expression of dominant negative Rac1 neither disrupts cell cell adhesion nor prevents assembly of new cadherin-based adhesion structures.

Antibodies↗

PubMed2000

An in vivo function for the transforming Myc protein: elicitation of the angiogenic phenotype.

The ability of neoplastic cells to recruit blood vasculature is crucial to their survival in the host organism. However, the evidence linking dominant oncogenes to the angiogenic switch remains incomplete. We demonstrate here that Myc, an oncoprotein implicated in many human malignancies, stimulates neovascularization. As an experimental model, we used Rat-1A fibroblasts that form vascular tumors upon transformation by Myc in immunocompromised mice. Our previous work and the use of neutralizing antibodies reveal that in these cells, the angiogenic switch is achieved via down-modulation of thrombospondin-1, a secreted inhibitor of angiogenesis, whereas the levels of vascular endothelial growth factor, a major activator of angiogenesis, remain high and unaffected by Myc. Consistent with this finding, overexpression of Myc confers upon the conditioned media the ability to promote migration of adjacent endothelial cells in vitro and corneal neovascularization in vivo. Furthermore, mobilization of estrogen-dependent Myc in vivo with the appropriate steroid provokes neovascularization of cell implants embedded in Matrigel. These data suggest that Myc is fully competent to trigger the angiogenic switch in vivo and that secondary events may not be required for neovascularization of Myc-induced tumors.

Animals↗

PubMed1999

Tracheal aspirate as a substrate for polymerase chain reaction detection of viral genome in childhood pneumonia and myocarditis.

BACKGROUND: Infectious respiratory disorders are important causes of childhood morbidity and mortality. Viral causes are common and may lead to rapid deterioration, requiring mechanical ventilation; myocardial dysfunction may accompany respiratory decompensation. The etiologic viral diagnosis may be difficult with classic methods. The purpose of this study was to evaluate polymerase chain reaction (PCR) as a diagnostic method for identification of causative agents. METHODS AND RESULTS: PCR was used to amplify sequences of viruses known to cause childhood viral pneumonia and myocarditis. Oligonucleotide primers were designed to amplify specific sequences of DNA virus (adenovirus, cytomegalovirus, herpes simplex virus, and Epstein-Barr virus) and RNA virus (enterovirus, respiratory syncytial virus, influenza A, and influenza B) genomes. Tracheal aspirate samples were obtained from 32 intubated patients and nucleic acid extracted before PCR. PCR results were compared with results of culture, serology, and antigen detection methods when available. In cases of myocarditis (n=7), endomyocardial biopsy samples were analyzed by PCR and compared with tracheal aspirate studies. PCR amplification of viral genome occurred in 18 of 32 samples (56%), with 3 samples PCR positive for 2 viral genomes. Amplified viral sequences included RSV (n=3), enterovirus (n=5), cytomegalovirus (n=4), adenovirus (n=3), herpes simplex virus (n=2), Epstein-Barr virus (n=1), influenza A (n=2), and influenza B (n=1). All 7 cases of myocarditis amplified the same viral genome from heart as found by tracheal aspirate. CONCLUSIONS: PCR is a rapid and sensitive diagnostic tool in cases of viral pneumonia with or without myocarditis, and tracheal aspirate appears to be excellent for analysis.

Adenoviridae Infections↗

PubMed1999

Spinal epidural hematoma associated with tissue plasminogen activator treatment of acute myocardial infarction.

We report a case of tissue plasminogen activator-associated spinal epidural hematoma in a patient who underwent treatment for myocardial infarction. Diagnostic magnetic resonance imaging was used within 24 hr of coronary artery stent implantation. We review the literature on thrombolytic-associated epidural spinal hematoma and discuss its management. Cathet. Cardiovasc. Intervent. 48:390-396, 1999.

Fibrinolytic Agents↗

PubMed1999

Volume (three-dimensional) fast spin-echo imaging of the lumbar spine.

RATIONALE AND OBJECTIVES: The purpose of this study was to prospectively evaluate a proton-density-weighted, three-dimensional (3D) volume fast spin-echo (SE) pulse sequence in the assessment of the lumbar spine for suspected spondylosis. MATERIALS AND METHODS: Twenty-eight patients referred for low back or lower extremity pain were imaged with both a two-dimensional (2D) protocol and a proton-density-weighted 3D volume fast SE imaging. The spinal canal, conus medullaris, intervertebral disks, neural foramina, bone marrow, and spinal alignment shown with the 3D volume fast SE pulse sequence were independently assessed by two neuroradiologists. These findings were compared with those of the routine 2D studies. RESULTS: Interpretation of disk protrusions and stenoses of the neural foramina were concordant between both protocols. No instance of cord abnormality was detected with either protocol. CONCLUSION: A 3D volume fast SE proton-density-weighted pulse sequence may provide information comparable to that of routine 2D imaging. Advantages of volume imaging include thinner sections, the capability of reconstruction into any plane, and the potential to decrease imaging time.

Adult↗

PubMed1999

Volume fast spin-echo imaging of the cervical spine.

RATIONALE AND OBJECTIVES: The authors prospectively evaluated a T2-weighted, three-dimensional (3D) volume, fast spin-echo (SE) pulse sequence in assessment of the cervical spine and compared it with standard imaging protocol. MATERIALS AND METHODS: Eighteen patients with neck pain underwent magnetic resonance (MR) imaging at 1.5 T with two-dimensional (2D) fast SE and axial 3D gradient-echo (GRE) protocols and with an additional sagittal T2-weighted volume fast SE protocol. The spinal cord and canal, neural foramina, and intervertebral disks were assessed by two neuroradiologists, and the results were compared with reports from the standard protocol. The quality of the partition (direct sagittal) and reconstructed images were evaluated. RESULTS: No differences existed in the assessment of spinal cord disease or disk herniation with 2D fast SE and volume fast SE imaging. Some mild variation occurred in assessment of the neural foramina. Partition images demonstrated a high level of resolution and contrast, while reconstructed images had consistently lower quality. However, this did not impede detection and grading of disk or spinal abnormalities, which were adequately shown on volume fast SE sagittal images. Neural foramina were well demonstrated on axial reconstructions from volume fast SE imaging. CONCLUSION: Volume fast SE imaging provides information about the spinal cord, canal, disks, and neural foramina that is comparable to the information provided by routine imaging. Its thinner sections and multiplanar reconstruction capability are advantages over 2D imaging. Its greater tissue contrast with better visualization of the cervical cord, greater signal-to-noise ratio, and less susceptibility artifact are advantages over 3D GRE imaging.

Adult↗

PubMed1999

Acquiring basic word order: evidence for data-driven learning of syntactic structure.

Recent studies indicate that young English-speaking children do not have a general understanding of the significance of SVO order in reversible sentences; that is, they seem to rely on verb-specific formulas (e.g. NPpusher-form of the verb PUSH-NPpushee) to interpret such sentences (Akhtar & Tomasello, 1997). This finding raises the possibility that young children may be open to learning non-SVO structures with novel transitive verbs. To test this hypothesis, 12 children in each of three age groups (two-year-olds, three-year-olds, and four-year-olds) were taught novel verbs, one in each of three sentence positions: medial (SVO), final (SOV), and initial (VSO). The younger age groups were equally likely to use the novel (non-English) orders spontaneously as to correct them to SVO order, whereas the oldest children consistently corrected these structures to SVO order. These results suggest that English-speaking children's acquisition of a truly general understanding of SVO order may be a gradual process involving generalization (learning) from examples. The findings are discussed in terms of recent data-driven learning accounts of grammar acquisition.

Child, Preschool↗

PubMed1999

Improved trigeminal and facial nerve tolerance following fractionated stereotactic radiotherapy for large acoustic neuromas.

The purpose of this study was to demonstrate improved cranial nerve tolerance following fractionated stereotactic radiotherapy for large acoustic neuromas, defined as tumours with pons-petrous distance (A) > 1 cm and midporous transverse diameter (A + Y) > 2 cm. Of 28 patients with acoustic neuromas treated with fractionated stereotactic radiotherapy, 19 had large tumours at high risk for radiosurgery-induced cranial neuropathy. Six patients received 36 Gy in six, weekly, fractions and 13 patients received 30 Gy in six, weekly, fractions. 15 patients had evaluable trigeminal nerve function and 16 had evaluable facial nerve function. At a median follow-up of 4.5 years, tumour shrinkage was seen in 10 patients and tumour size was stable in nine. None of the patients developed any evidence of transient, or permanent, trigeminal or facial neuropathy at any time during their follow-up period. Fractionated stereotactic radiotherapy may offer a superior therapeutic ratio to single fraction stereotactic radiosurgery in the management of large acoustic neuromas, as evidenced by the absence of post-treatment trigeminal and facial neuropathy.

Adult↗

PubMed1999

Acute hyperthermia following stereotactic radiosurgery for pituitary adenoma.

A 71-year-old male presented with a large pituitary adenoma with superior extension into the optic chiasm and suprasellar cistern. He was treated with stereotactic radiosurgery to a dose of 16 Gy. Approximately 1 h after radiosurgery he developed fever; his temperature peaked at 105.1 degrees F and normalized about 20 h later. This case demonstrates that acute hyperthermia is a potential complication following high dose stereotactic radiosurgery for large pituitary tumours.

Acute Disease↗

PubMed1998

The Saccharomyces cerevisiae SOP1 and SOP2 genes, which act in cation homeostasis, can be functionally substituted by the Drosophila lethal(2)giant larvae tumor suppressor gene.

By complementation of a salt-sensitive mutant of Saccharomyces cerevisiae, we cloned the SOP1 gene, encoding a 114.5-kDa protein of 1033 amino acids. Cells deleted for SOP1 exhibited sensitivity to sodium stress, but showed no sensitivity to general osmotic stress. Following exposure of sop1Delta cells to NaCl stress, the intracellular Na+ level and the Na+/K+ ratio rose to values significantly higher than in wild type cells. Deletion of SOP2, encoding a protein sharing 54% amino acid identity with Sop1p, produced only slight Na+ sensitivity. Cells carrying a sop1Deltasop2Delta double deletion became, however, hypersensitive to Na+ and exhibited increased sensitivity also to Li+ and K+, suggesting involvement of both SOP1 and SOP2 in cation homeostasis. The predicted amino acid sequences of Sop1p and Sop2p show significant homologies with the cytoskeletal-associated protein encoded by the Drosophila lethal(2)giant larvae tumor suppressor gene. Immunolocalization of Sop1p revealed a cytoplasmic distribution and cell fractionation studies showed that a significant fraction of Sop1p was recovered in a sedimentable fraction of the cytosolic material. Expression of a Drosophila l(2)gl cDNA in the sop1Deltasop2Delta strain partially restored the Na+ tolerance of the cells, indicating a functional relationship between the Sop proteins and the tumor suppressor protein, and a novel function in cell homeostasis for this family of proteins extending from yeast to human.

Adaptor Proteins, Signal Transducing↗