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Biomedical subjects

S B Lee

Publications and source records attributed to S B Lee.

At least 55 records · Page 3Linked to original sources

Analysis of growth rate effects on productivity of recombinant Escherichia coli populations using molecular mechanism models. Reprinted from Biotechnology and Bioengineering, Vol. 26, Issue 1, Pages 66-73 (1984).

The influence of growth rate on Escherichia coli plasmid content and expression of a cloned-gene product has been described by a mathematical model based upon the molecular mechanism of lambdadv plasmid replication and known relationships between growth rate and transcription and translation activities of the host cell. The model simulates correctly decreases in plasmid content with increasing growth rate as observed experimentally for pBR322, NR1, R1, and Col E1 plasmids. A maximum with respect to growth rate in intracellular product accumulation is indicated by the model, as is a transient overshoot in product concentration following a shift from smaller to larger growth rate. Available data, although very limited, show the same trends. These results, obtained without parameter or kinetic form adjustments or manipulation, clearly illustrate the advantages of kinetic descriptions of recombinant systems based upon the pertinent molecular mechanisms.

Escherichia coli↗

Upregulation of phospholipase D in astrocytes in response to transient forebrain ischemia.

Previous in vitro studies using cell cultures or brain slices have demonstrated that phospholipase D (PLD) in the nervous system is involved in the signaling mechanism in response to a variety of agonists. However, little is known about the pathophysiological role of PLD-mediated signaling in the adult brain. We examined the changes in the expression of a PLD isozyme, PLD1, in the adult rat hippocampus, using immunological approaches and an assay for PLD activity after transient forebrain ischemia (four-vessel occlusion model) that results in the selective delayed death of CA1 pyramidal cells and induces reactive astrocytes in the CA1 subfield. In the control hippocampus, PLD1 the level of immunoreactivity was very low. After ischemia, in parallel with the results of Western blot analysis and the PLD activity assay, immunohistochemical analysis of PLD1 demonstrated that the immunoreactive proteins peaked at 7-14 days and were most prominent in the CA1 and the dentate hilar region. The temporal and spatial patterns of immunoreactivity of both PLD1 and glial fibrillary acidic protein (GFAP) were very similar, indicating that reactive astrocytes express PLD1, confirmed by double staining for PLD1 and GFAP. These results demonstrate that reactive astrocytes upregulate PLD in vivo after injury in the adult rat hippocampus.

Animals↗

Acidosis induces necrosis and apoptosis of cultured hippocampal neurons.

Acidosis, hypoxia, and hypoglycemia rapidly and transiently appear after reduction of cerebral blood flow. Acidosis also accompanies head trauma and subarachnoid hemorrhage. These insults result in necrotic and apoptotic loss of neurons. We previously demonstrated that transient acidification of intracellular pH from 7.3 to 6.5 induces delayed neuronal loss in cultured hippocampal slices (49). We now report that acidosis induced both necrotic and apoptotic loss of neurons. Necrosis and apoptosis were distinguished temporally and pharmacologically. Necrosis appeared rapidly and was dose dependent with the duration of the acidosis treatment. Apoptosis was delayed with maximal number of apoptotic cells seen with a 30-min acidosis treatment. Apoptotic neuronal loss was accompanied by DNA fragmentation and was blocked by inhibitors of protein and RNA synthesis, ectopic expression of the anti-apoptotic gene bcl-2, or an inhibitor of caspases, proteases known to be activated during apoptosis. Necrotic neuronal loss was unaffected by these treatments. Hypothermia, a treatment known to attenuate neuronal loss following a variety of insults, blocked both acidosis-induced necrosis and apoptosis. These results indicate that acidosis is neurotoxic in vitro and suggest that acidosis contributes to both necrotic and apoptotic neuronal loss in vivo.

Acidosis↗

Synthesis and biological activity of aspirin derivatives.

Aspirin has been widely used as analgesic and anti-inflammatory drug. Recently, it was elucidated that aspirin have anti-coaggregatory effect in low dose. This study was carried out to investigate the synthesis of aspirin derivatives from aspirin and aromatic compound of antioxidant and its biological activities. Synthesis of aspirin derivatives was prepared by esterification in the presence of 1,1-carbonyldiimidazole. Biological activities was examined using effect of anti-coagulant on bleeding time, effect of antioxidant and effect of anti-platelet aggregation. As a result, SJ-101 showed strong antioxidative activity and anti-coagulant activity among four compounds. Anti-platelet aggregation of SJ-101 was examined by collagen, ADP, PAF method. SJ-101 exhibited more stronger activity to aspirin at collagen aggregation reaction. These finding demonstrates that SJ-101 is useful as care drug of aging and old-disease because of its has antioxidant activity, anti-coagulant activity and anti-platelet activity.

Animals↗

Insertion of a retroviral solo long terminal repeat in mdr-3 locus disrupts mRNA splicing in mice.

Previously, the abermectin-induced neurotoxicity of subpopulation of CF-1 mice was shown to be caused by the deficiency of mdr-3 P-glycoprotein. Here, we have characterized the molecular nature of the mdr-3 gene mutation in this subpopulation of CF-1 mice. The size of mdr-3 mRNA transcript from ivermectin-sensitive mutant mice was different from that of wild-type mice. Sequence analysis of RT-PCR products isolated from the mutant brain disclosed that the exon 23 of the mdr-3 gene is deleted or altered in the transcripts. The analysis of the genomic locus revealed an insertion of a solo long terminal repeat (LTR) of the ecotropic murine leukemia virus in the reverse orientation in the intron of the mdr-3 gene, causing abnormal splicing and thereby disrupting the mdr-3 gene function. In addition, histopathological analysis of the brains of the ivermectin-treated mutants revealed selective neuronal degeneration in the hippocampal CA3 region. This is the first reported case of a gene mutation induced by a solo retroviral LTR with a phenotypic consequence in the mouse, and may provide new insights into the understanding of the effects of viral solo LTR sequences on mammalian gene expression.

ATP Binding Cassette Transporter, Subfamily B↗

Identity of mouse IA-2 and PTP35 genes of the tyrosine phosphatase family, and their expression in neuroendocrine tissues.

Recently, IA-2, one of the major diabetic autoantigens, and PTP35 cDNA were independently isolated by subtraction cloning using insulinoma cells and a polymerase chain reaction (PCR)-based search for conserved sequences using NIH3T3 fibroblast cell line, respectively. By Southern blot analysis and nucleotide sequence determination of reverse transcription PCR products, we showed that IA-2 and PTP35 are identical and exist as a single gene in a mouse genome. The expression of IA-2/PTP35 messages was detected by northern blot analysis in MIN6N8 cells, an insulinoma cell line derived from non-obese diabetic mice, but its expression level was not affected by the ambient glucose level, phorbol-12-myristate 13-acetate or tumour necrosis factor-alpha. We also generated polyclonal antibodies to murine IA-2/PTP35 by immunization with recombinant proteins. Subsequent immunohistochemical analysis using these polyclonal antibodies disclosed that IA-2/PTP35 is strongly expressed in mouse neuroendocrine tissues such as pancreatic islets and the hypothalamus-pituitary gland. These results suggest that IA-2/PTP35 functions primarily in neuroendocrine tissues.

3T3 Cells↗

The bursal and articular sides of the supraspinatus tendon have a different compressive stiffness.

OBJECTIVE: To measure the compressive stiffness of the supraspinatus tendon and to determine whether regional difference exists in the bursal and articular side of the tendon. DESIGN: Indentation testing was performed on both the bursal and articular sides of the supraspinatus tendon, focused on the 'critical area', where rotator cuff tears often occur. BACKGROUND: When the supraspinatus tendon wraps around the humeral head or is under impingement condition, compressive force on the tendon surface is expected. Therefore, compressive stress has been recently considered to be one of the important factors associated with the cuff tear. The mechanical properties would be essential for analytic modeling of stress distribution. METHODS: Indentation tests were performed at 15 locations on the bursal and articular surfaces of the supraspinatus tendon. A mathematical model with exponential relationship was used to describe the measured force-deformation relationship and to calculate the compressive stiffness of the supraspinatus tendon. RESULTS: The over-all initial stiffness on the bursal and articular sides of the tendon was significantly different. On the bursal side, the anterior third had a significantly higher initial stiffness than the other thirds on average. On the articular side, initial stiffness at location 10 mm proximal to the greater tuberosity was significantly higher than the rest on average. CONCLUSIONS: The compressive stiffness of the supraspinatus tendon was found to be non-homogenous throughout the structure.RelevanceNon-homogenous compressive stiffness of the supraspinatus tendon would affect the load transmission within the tendon, which might be associated with the potential mechanism of tear. Such characteristics needs to be considered when performing finite element modeling of stress fields in the tendon.

Aged↗

Effects of exogenous compatible solutes on growth of the hyperthermophilic archaeon Sulfolobus solfataricus.

Six known compatible solutes as well as twenty L-amino acids were individually added to a glucose minimal medium and their effects on the growth of Sulfolobus solfataricus (DSM 1617) were examined. Among the compatible solutes tested, putrescine, trehalose, and l-glutamate enhanced the growth of S. solfataricus. On the other hand, glycine betaine, choline, and L-proline showed little or no influence on cell growth. When cells were grown in the glucose medium supplemented with trehalose or L-glutamate, S. solfataricus preferentially utilized the compatible solute over glucose. The growth-enhancement effect of L-glutamate was also observed to be dependent on the glucose concentration in the medium: growth enhancement was higher when the concentration of glucose was low and gradually decreased with increasing glucose concentration. Interestingly, the effects of amino acids on cell growth differed markedly depending on the chemical nature of the amino acid added. While acidic amino acids-L-glutamate and L-aspartate-enhanced the growth rate, almost no growth was observed in the presence of glycine, L-leucine, L-valine, L-phenylalanine, L-threonine, L-methionine, or L-cysteine. Among all the low-molecular-weight solutes tested in this study, the growth-stimulation effect was most profound in the presence of L-glutamate. When S. solfataricus cells were grown in a glucose (1.0 g/l) medium supplemented with 3.0 g/l L-glutamate, the maximal cell density and growth rate were about 3.2- and 2.3-fold higher than those obtained without L-glutamate.

Journal Article↗

Effects of hen egg yolk immunoglobulin in passive protection of rainbow trout against Yersinia ruckeri.

Anti-Yersinia ruckeri egg yolk immunoglobulin (IgY) was transferred to egg yolk after immunization of White Leghorn hens with formalin-killed whole cells of serovar 1 (RS1154) and serovar 2 (RS1153)Y. ruckeri and its lipopolysaccharide (LPS). The IgY was specific for its homologous LPS in western immunoblot, whereas some protein bands were commonly recognized, even by IgY from eggs of unimmunized hens. Purified LPS from both Y. ruckeri serovar types 1 and 2 had a very poor immunogenicity. The IgY activity was stable when processed into pellet form by a microbial transglutaminase treatment and showed a considerable resistance against acid pepsin for at least 2 h. Feeding specific anti-serovar 1 Y. ruckeri IgY to fish either before or after immersion infection produced marginal reductions in mortalities and in intestine infection. The same IgY did passively protect rainbow trout against infection when administered by intraperitoneal injection 4 h before an immersion challenge.

Animals↗

Transgenic rice plants expressing a Bacillus subtilis protoporphyrinogen oxidase gene are resistant to diphenyl ether herbicide oxyfluorfen.

Protoporphyrinogen oxidase (Protox), the penultimate step enzyme of the branch point for the biosynthetic pathway of Chl and hemes, is the target site of action of diphenyl ether (DPE) herbicides. However, Bacillus subtilis Protox is known to be resistant to the herbicides. In order to develop the herbicide-resistant plants, the transgenic rice plants were generated via expression of B. subtilis Protox gene under ubiquitin promoter targeted to the cytoplasm or to the plastid using Agrobacterium-mediated gene transformation. The integration and expression of the transgene were investigated at T0 generation by DNA and RNA blots. Most transgenic rice plants revealed one copy transgene insertion into the rice genome, but some with 3 copies. The expression levels of B. subtilis Protox mRNA appeared to correlate with the copy number. Furthermore, the plastidal transgenic lines exhibited much higher expression of the Protox mRNA than the cytoplasmic transgenic lines. The transgenic plants expressing the B. subtilis Protox gene at T0 generation were found to be resistant to oxyfluorfen when judged by cellular damage with respect to cellular leakage, Chl loss, and lipid peroxidation. The transgenic rice plants targeted to the plastid exhibited higher resistance to the herbicide than the transgenic plants targeted to the cytoplasm. In addition, possible resistance mechanisms in the transgenic plants to DPE herbicides are discussed.

Agrobacterium tumefaciens↗

Eye cancer incidence in Singapore.

AIM: To describe the epidemiological characteristics of patients with primary intraocular, conjunctival, and orbital cancers seen in Singapore from 1968 to 1995. METHODS: Epidemiological data of all cancers diagnosed in Singapore are collected by the Singapore Cancer Registry. The data of all cases of Singapore residents with eye cancers (ICD-9, site 190) were retrieved for analysis. This includes intraocular, conjunctival, and orbital cancers but excludes cancer of the eyelids. RESULTS: There were 125 patients of which 67 (53.6%) were male and 58 (46.4%) were female. The average annual age standardised incidences for male and female Singapore residents were 1.89 and 1.81 per million respectively. The most common cancer was retinoblastoma (53.6%), followed by malignant melanoma (19.2%) and squamous cell carcinoma (11.2%). The most common cancer among patients younger than 15 years was retinoblastoma (95.7%) and that for those 15 years and older was malignant melanoma (42.6%). The most common subsite was the retina (53.6%), followed by conjunctiva (12.8%), orbit (8.8%), and lacrimal gland (6.4%). CONCLUSION: The annual age standardised incidence have been stable for the 28 years studied. Retinoblastoma is much more common than melanoma in Singapore. These expanded epidemiological characteristics serve to provide ophthalmologists and epidemiologists with a foundation to monitor future disease patterns in Singapore and provide a basis for comparison with other selected populations elsewhere.

Adolescent↗

Plasma cell polyp of the vocal fold.

Plasma cell polyps of the vocal fold (plasma cell granulomas) are rare inflammatory polyps of the larynx. They should be included in the clinical and histological differential diagnosis of laryngeal polyps. Histologically they are polyclonal aggregates of plasma cells. It is essential to distinguish them from monoclonal, neoplastic plasma cell proliferations. The treatment of choice is surgical resection, although radiotherapy, laser ablation, antibiotics and steroids have been used successfully. We present a case of plasma cell granuloma presenting as a vocal fold polyp, treated surgically.

Aged↗

The effect of a glenoid defect on anteroinferior stability of the shoulder after Bankart repair: a cadaveric study.

BACKGROUND: An osseous defect of the glenoid rim is sometimes caused by multiple recurrent dislocations of the shoulder. It is generally thought that a large defect should be treated with bone-grafting, but there is a lack of consensus with regard to how large a defect must be in order to necessitate this procedure. Some investigators have proposed that a defect must involve at least one-third of the glenoid surface in order to necessitate bone-grafting. However, it is difficult to determine (1) whether a defect involves one-third of the glenoid surface and (2) whether a defect of this size is critical to the stability of the shoulder after a Bankart repair. The purposes of the present study were (1) to create and quantify various sizes of osseous defects of the glenoid and (2) to determine the effect of such defects on the stability and motion of the shoulder after Bankart repair. METHODS: The glenoids from sixteen dried scapulae were photographed, and the images were scanned into a computer. The average shape of the glenoid was determined on the basis of the scans, and this information was used to design custom templates for the purpose of creating various sizes of osseous defects. Ten fresh-frozen cadaveric shoulders then were obtained from individuals who had been an average of seventy-nine years old at the time of death, and all muscles were removed to expose the joint capsule. With use of a custom multiaxis electromechanical testing machine with a six-degrees-of-freedom load-cell, the humeral head was translated ten millimeters in the anteroinferior direction with the arm in abduction and external rotation as well as in abduction and internal rotation. With a fifty-newton axial force constantly applied to the humerus in order to keep the humeral head centered in the glenoid fossa, the peak force that was needed to translate the humeral head a normalized distance was determined under eleven sequential conditions: (1) with the capsule intact, (2) after the creation of a simulated Bankart lesion, (3) after the capsule was repaired, (4) after the creation of an anteroinferior osseous defect with a width that was 9 percent of the glenoid length (average width, 2.8 millimeters), (5) after the capsule was repaired, (6) after the creation of an osseous defect with a width that was 21 percent of the glenoid length (average width, 6.8 millimeters), (7) after the capsule was repaired, (8) after the creation of an osseous defect with a width that was 34 percent of the glenoid length (average width, 10.8 millimeters), (9) after the capsule was repaired, (10) after the creation of an osseous defect with a width that was 46 percent of the glenoid length (average width, 14.8 millimeters), and (11) after the capsule was repaired. RESULTS: With the arm in abduction and external rotation, the stability of the shoulder after Bankart repair did not change significantly regardless of the size of the osseous defect (p = 0.106). With the arm in abduction and internal rotation, the stability decreased significantly as the size of the osseous defect increased (p<0.0001): the translation force in shoulders in which the width of the osseous defect was at least 21 percent of the glenoid length (average width, 6.8 millimeters) was significantly smaller than the force in shoulders without an osseous defect. The range of external rotation in shoulders in which the width of the osseous defect was at least 21 percent of the glenoid length was significantly less than that in shoulders without a defect (p<0.0001) because of the pretensioning of the capsule caused by closing the gap between the detached capsule and the glenoid rim. The average loss of external rotation was 25 degrees per centimeter of defect. CONCLUSIONS: An osseous defect with a width that is at least 21 percent of the glenoid length may cause instability and limit the range of motion of the shoulder after Bankart repair.

Aged↗

Dynamic glenohumeral stability provided by the rotator cuff muscles in the mid-range and end-range of motion. A study in cadavera.

BACKGROUND: Both static and dynamic factors are responsible for glenohumeral joint stability. We hypothesized that dynamic factors could potentially operate throughout the entire range of glenohumeral motion, although capsuloligamentous restraints (a static factor) have been thought to be primarily responsible for stability in the end-range of motion. The purpose of this study was to quantitatively compare the dynamic glenohumeral joint stability in the end-range of motion (the position of anterior instability) with that in the mid-range by investigating the force components generated by the rotator cuff muscles. METHODS: Ten fresh-frozen shoulders from human cadavera were obtained, and all soft tissues except the rotator cuff were removed. The glenohumeral capsule was resected after the rotator cuff muscles had been released from the scapula. A specially designed frame positioned the humerus in 60 degrees of abduction and 45 degrees of extension with respect to the scapula. The compressive and shear components on the glenoid were measured before and after a constant force was applied individually to each muscle with the humerus in five different positions (from neutral to 90 degrees of external rotation). The dynamic stability index, a new biomechanical parameter reflecting these force components and the concavity-compression mechanism, was calculated. The higher the dynamic stability index, the greater the dynamic glenohumeral stability. RESULTS: In the mid-range of motion, the supraspinatus and subscapularis provided higher dynamic stability indices than did the other muscles (p < 0.05). On the other hand, when the position of anterior instability was simulated in the end-range of motion, the subscapularis, infraspinatus, and teres minor provided significantly higher dynamic stability indices than did the supraspinatus (p < 0.005). CONCLUSIONS: The rotator cuff provided substantial anterior dynamic stability to the glenohumeral joint in the end-range of motion as well as in the mid-range. CLINICAL RELEVANCE: A glenohumeral joint with a lax capsule and ligaments might be stabilized dynamically in the end-range of motion if the glenoid concavity is maintained and the function of the external and internal rotators, which are efficient stabilizers in this position, is enhanced.

Aged↗

A comparison of Min's glasses and conventional occlusion therapy in the treatment of amblyopic children: a prospective study.

Min's glasses are specially manufactured in order to enhance the satisfaction of the wearer and increase treatment effectiveness. We compared the effectiveness of Min's glasses with that of conventional occlusion therapy in amblyopic pediatric patients. We prospectively studied 60 amblyopic patients. For four to 30 months, 24 of the patients were treated with conventional patch occlusion (patch group) and 36 of them were treated with Min's glasses (glasses group). We compared the improvement in visual acuity and the treatment compliance between both groups, according to age (over six and less than six), sex, type of amblyopia, and the duration of treatment. Sixteen patients (66.7%) in the patch group and 32 patients (88.9%) in glasses group showed improvement in visual acuity (p = 0.002). The pre-treatment average log MAR acuity was 0.56 +/- 0.25 in the patch group, and 0.59 +/- 0.25 in the glasses group. The average improvement in visual acuity was a 0.17 log MAR score in the patch group, and a 0.31 log MAR score in the glasses group (p = 0.004). Compliance was 59.54% in the patch group and 83.44% in the glasses group (p = 0.012). The pre-treatment average log MAR acuity was 0.49 +/- 0.23 for children over six years of age in the patch group, and 0.58 +/- 0.28 for children over six years of age in the glasses group. For children over six years of age in the glasses group the improvement in visual acuity (0.29 log MAR score) was greater than for children over six years of age in the patch group (0.06 log MAR score) (p = 0.0003). The pre-treatment average log MAR acuity was 0.55 +/- 0.22 for female patients in the patch group, and 0.60 +/- 0.25 for female patients in the glasses group. Female patients in the glasses group also showed a greater visual acuity improvement (0.29 log MAR score) than female patients in the patch group (0.14 log MAR score) (p = 0.0028). However, there were no differences between the groups in patients less than six years of age and in male patients. In conclusion, Min's glasses were more effective than conventional treatment with a patch in improving visual acuity and encouraging compliance in pediatric amblyopic patients, especially in children over six years of age and in girls.

Amblyopia↗

Suppression of TGF-beta signaling in both normal conjunctival fibroblasts and pterygial body fibroblasts by amniotic membrane.

PURPOSE: When used as an alternative substrate following bare sclera removal of pterygium and other ocular surface diseases, amniotic membrane transplantation can reduce scarring on the reconstructed conjunctival surface. This study was carried out to determine if the amniotic membrane (AM) suppresses the expression of the TGFb signaling system in cultured normal conjunctival (HCF) and pterygial body fibroblasts (PBF). METHODS: HCF and PBF were cultured on AM and plastic wells in serum-containing and serum-free DMEM with or without TGF-beta1. Total RNA was extracted and subjected to Northern hybridization with probes of TGF-beta1, b2 and b3; TGF-beta receptors (TGF- beta R) type I, II and III; a-smooth muscle actin (alpha-SM), b1-integrin, CD44, fibroblast growth factor receptor 1 (FGF-R1/ flg) and platelet-derived growth factor receptor b (PDGFR-beta); and GAPDH as a loading control. MTT assay was used for cell proliferation. RESULTS: Amniotic membrane markedly suppressed the transcript expression of TGF-beta2, b3 and all three types of TGF-beta receptors by both fibroblasts as compared to their cultures on plastic surface. In addition, expression of CD44 transcript was also markedly suppressed while that of b1 integrin, a-SM actin, and FGFR1/flg was mildly suppressed. In contrast, expression of TGF-beta1 and PDGFR-beta remained largely unchanged. The cell proliferation of HCF and PBF grown on AM was also significantly suppressed. CONCLUSIONS: Amniotic membrane matrix uniquely suppresses TGF- beta signaling in both types of fibroblasts. It may also suppress signaling via CD44, b1 integrin and FGFR1/flg. As a result, the phenotype may become less mitogenic, contractile and fibrogenic. These data support in part why amniotic membrane transplantation has an anti-scarring effect for conjunctival surface reconstruction.

Actins↗

Regulation of collagenase, stromelysin, and urokinase-type plasminogen activator in primary pterygium body fibroblasts by inflammatory cytokines.

PURPOSE: To examine the expression patterns of extracellular matrix degrading enzymes in cultured primary pterygium body fibroblasts activated by cytokines and growth factors potentially derived from ocular surface epithelial cells and tears. METHODS: EGF, TGF-alpha, PDGF-BB, IL-1beta, bFGF, TGF-beta1, TNF-alpha, or IL-6 were added at 10 ng/ml to early passaged primary pterygium body fibroblasts (PBF) or normal human conjunctival fibroblasts (HJF) in a serum-free medium. Expression of transcripts and proteins of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, and uPA was determined by Northern hybridization, ELISA, and Western blotting, respectively. Gelatin and casein zymographies were performed in their serum-free conditioned media with or without enzyme inhibitors to determine the activity of MMP-2 and -3, respectively. RESULTS: IL-1beta and TNF-alpha dramatically increased the mRNA and protein expression of MMP-1 and MMP-3 in cultured PBF when compared to normal HJF and to their nonstimulated counterparts cultured in a serum-free medium. EGF and TGF-alpha also upregulated MMP-3 in PBF when compared to HJF. The transcript levels of MMP-2 were high but stable for the two cell types regardless of the cytokine treatment. Both TIMP-1 and TIMP-2 expressions were not influenced by the cell type or the cytokine treatment. MMP-9 was not expressed in either of these two types of fibroblasts. Both IL-1beta and TNF-alpha induced a significant decrease in uPA expression in PBF, whereas bFGF induced a slight increase in both HJF and PBF. CONCLUSIONS: Chronic inflammatory stimulation by IL-1beta and TNF-alpha, which potentially can be derived from the ocular surface and tears, may be responsible for increased expression of MMPs in cultured PBF. These data have clinical implications on progression of pterygium and recurrence associated with incomplete excision of primary PBF under the influence of ocular surface inflammation. Suppression of intraoperative and postoperative inflammation may be a new strategy to prevent pterygium recurrence.

Blotting, Northern↗

Ablation of P/Q-type Ca(2+) channel currents, altered synaptic transmission, and progressive ataxia in mice lacking the alpha(1A)-subunit.

The Ca(2+) channel alpha(1A)-subunit is a voltage-gated, pore-forming membrane protein positioned at the intersection of two important lines of research: one exploring the diversity of Ca(2+) channels and their physiological roles, and the other pursuing mechanisms of ataxia, dystonia, epilepsy, and migraine. alpha(1A)-Subunits are thought to support both P- and Q-type Ca(2+) channel currents, but the most direct test, a null mutant, has not been described, nor is it known which changes in neurotransmission might arise from elimination of the predominant Ca(2+) delivery system at excitatory nerve terminals. We generated alpha(1A)-deficient mice (alpha(1A)(-/-)) and found that they developed a rapidly progressive neurological deficit with specific characteristics of ataxia and dystonia before dying approximately 3-4 weeks after birth. P-type currents in Purkinje neurons and P- and Q-type currents in cerebellar granule cells were eliminated completely whereas other Ca(2+) channel types, including those involved in triggering transmitter release, also underwent concomitant changes in density. Synaptic transmission in alpha(1A)(-/-) hippocampal slices persisted despite the lack of P/Q-type channels but showed enhanced reliance on N-type and R-type Ca(2+) entry. The alpha(1A)(-/-) mice provide a starting point for unraveling neuropathological mechanisms of human diseases generated by mutations in alpha(1A).

Animals↗