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Biomedical subjects

J W Chandler

Publications and source records attributed to J W Chandler.

At least 19 recordsLinked to original sources

The effect of minoxidil on keratocyte proliferation in cell culture.

PURPOSE: To determine if minoxidil inhibits keratocyte proliferation in a nontoxic manner. METHODS: Rabbit keratocytes were cultured in Eagle's minimum essential medium supplemented with fetal bovine serum. Minoxidil varying in concentration from 10(0) to 10(3) micrograms/ml was added to the culture medium and incubated for 7 days. The cultures were inspected for morphologic appearance and the cell number was determined at 1, 3 and 7 days after the addition of minoxidil. After 7 days of incubation, minoxidil was withdrawn from the cell culture medium and the cells were examined 3 and 7 days thereafter. In addition, a nonradioactive cytotoxic assay was performed to determine if toxicity is associated with the presence of minoxidil. RESULTS: Minoxidil inhibited keratocyte proliferation in a dose-dependent fashion. 29% of control growth was achieved when keratocytes were cultured for 7 days in 10(3) micrograms/ml, whereas 82% control growth was achieved when keratocytes were cultured in 10(2) micrograms/ml of minoxidil. Intermediate concentrations between 10(2) and 10(3) micrograms/ml produced a linear decline in cell counts in a dose-dependent fashion. The concentration of minoxidil required for 50% control growth at 7 days extrapolated from the dose-response curve was 600 micrograms/ml. Upon withdrawal of minoxidil, cell counts returned to baseline for concentrations of 10(2) micrograms/ml or less. Phase contrast microscopy revealed that the presence of minoxidil was associated with intercellular separation, enlargement of cell bodies and elongated processes. After the withdrawal of minoxidil, the cells in all media reassumed the morphological features of normal keratocytes which included a regular fusiform shape and extensive intercellular contact. The nonradioactive cytotoxic assay revealed the lack of cytotoxicity at all concentrations of minoxidil based on a lack of lactate dehydrogenase release. CONCLUSIONS: Minoxidil inhibits keratocyte proliferation by a nontoxic mechanism. It might be particularly useful for modulating corneal wound healing following excimer laser photorefractive keratectomy.

Animals↗

Structure and function of the vp1 gene homologue from the resurrection plant Craterostigma plantagineum Hochst.

A gene that is homologous to the vp1 genes from maize and rice, the Arabidopsis abi3 gene and PvAlf from Phaseolus vulgaris was isolated from the resurrection plant, Craterostigma plantagineum Hochst, The C. plantagineum gene (epvp1) encodes a protein of 688 amino acids and contains five introns at positions identical to those in the Arabidopsis, maize and rice homologues. The cpvp1 transcript is present in mature seeds and in young seedlings up to 8 days following germination. The ability of the cpvp1 gene product to activate target genes was demonstrated by transient expression experiments in tobacco protoplasts using reporter gene constructs containing the beta-glucuronidase (GUS) gene fused to the promoter of two late embryogenesis abundant (LEA)-like genes, CDeT27-45 and CDeT6-19 from C. plantagineum, or the promoter of a maize gene encoding a 22-kDa zein seed storage protein.

Abscisic Acid↗

Analysis of cDNA clones encoding sucrose-phosphate synthase in relation to sugar interconversions associated with dehydration in the resurrection plant Craterostigma plantagineum Hochst.

Sucrose-phosphate synthase (SPS) is a key enzyme in the regulation of sucrose metabolism, being responsible for the synthesis of sucrose 6-phosphate from fructose 6-phosphate and uridine 5'-diphosphate-glucose. We report on the isolation and characterization of cDNA clones encoding SPS from Craterostigma plantagineum Hochst., a resurrection plant in which the accumulation of sucrose is considered to play an important role in tolerance to severe protoplastic dehydration. Two distinct classes of cDNAs encoding SPS were isolated from C. plantagineum, and are represented by the clones Cpsps1 and Cpsps2. The transcripts corresponding to both cDNAs decrease to very low levels in dehydrating leaves of C. plantagineum. Only the Cpsps1 transcript occurs in the roots, where it is present at a higher level than in leaves and increases upon dehydration of the plant. Higher enzymatic activities have been determined in protein extracts of dehydrated tissues compared with untreated tissues, which correlates with an increase in protein levels. It is suggested that the overall regulation of SPS is strongly influenced by the changing composition of the cytoplasm in C. plantagineum leaves during the dehydration-rehydration cycle.

Amino Acid Sequence↗

Polyhexamethylene biguanide (PHMB) in the treatment of experimental Fusarium keratomycosis.

PURPOSE: We wanted to determine whether topical polyhexamethylene biguanide (PHMB) 0.02% was effective in the treatment of experimental Fusarium keratomycosis in rabbits. METHODS: Fusarium solani keratomycosis was induced in the eyes of 12 New Zealand white rabbits. The rabbits were treated with PHMB 0.02% in one eye and placebo in the other eye for 6 days. The rabbits were evaluated in a masked fashion using a standardized system for clinical progression of the disease. Then the corneas were trephined and growth of F. solani in colony-forming units per milliliter (CFU/ml) determined. RESULTS: Clinical evaluation demonstrated no significant mean difference (p > 0.10) in clinical scores between treated and control eyes on day 6 (0.583 +/- 2.503). There was a significant mean CFU difference (p = 0.06) between treated eyes and control eyes (182.5 +/- 314.44). Seven of 12 eyes (58%) in the PHMB group exhibited no growth, whereas two of 12 (17%) eyes reported no growth in the control group. One of 12 eyes (8%) reported > 100 CFU in the PHMB group, whereas seven of 12 eyes (58%) reported > 100 CFU in the control group. CONCLUSIONS: PHMB 0.02% was effective in significantly reducing the fungal growth in our rabbit model of Fusarium keratomycosis. The future role of PHMB in the treatment of Fusarium keratitis needs to be further evaluated.

Administration, Topical↗

Localization of endothelin-like immunoreactivity and endothelin binding sites in rabbit eyes.

Immunohistochemistry and in vitro autoradiographic techniques were used to investigate the localization of endothelin (ET)-like immunoreactivity and [125I]ET-1 binding sites in sections of ocular tissues of normal rabbits. ET-like immunoreactivity was localized to the corneal endothelium and the pre-epithelial layer of the cornea, which was specifically inhibited by addition of exogenous ET-1. Immunoreactivity for ET was also observed in the retinal ganglion layer and periocular fat tissues. These, however, were not inhibited in the presence of ET-1. Autoradiography showed binding sites for ET-1 in the iris and ciliary body. Emulsion-dipped slides of sections of rabbit eyes incubated with [125I]ET-1 showed specific labeling for binding sites over structures that were identified from serial sections stained with hematoxylineosin. These were the anterior stroma of the iris root, the walls of capillaries, the circular arteries, and the veins of the ciliary body and extraocular muscles. No binding sites were found in the cornea or conjunctiva. The difference in regional distributions of ET storage and binding sites in rabbit ocular tissue suggests that the peptide may have specific physiologic functions.

Animals↗

Dynamic shifts in corneal topography after radial and transverse keratotomy.

PURPOSE: The authors aimed to quantitate the dynamic patterns of change in corneal topography after multistaged radial and transverse keratotomy using digitized video-keratography. METHODS: Single and paired radial and transverse keratotomies, with videokeratoscopy between each stage and at the end of the procedure, were performed on fresh animal cadaver eyes using an artificial orbit system. RESULTS: All incisions led to central flattening. A single radial keratotomy caused flattening adjacent to the incision, and steepening 180 degrees away. A paired radial keratotomy caused increased flattening in the meridian of the incisions, and less flattening 90 degrees away. A single transverse incision caused steepening adjacent to the incision and diffuse flattening elsewhere. A paired transverse incision caused flattening near the optical center along the meridian bisecting the incisions and steepening 90 degrees away. CONCLUSION: The authors have demonstrated that computerized videokeratography can be used successfully to systematically quantitate dioptric shifts in multiple hemimeridians and measurement zone diameters after refractive surgery.

Animals↗

Localization and characterization of binding sites for endothelin in Harder's gland in rabbits.

The characterization and localization of binding sites for endothelin-1 (ET-1) labeled with iodine 125I were investigated in homogenized tissues and sections of Harder's glands of normal rabbits. The membrane of Harder's glands was harvested and incubated with 125I-ET-1 (0.25-1 nmol/L) in 20 +/- 4 mg of protein per 0.25 mL at 37 degrees C for 90 min in the presence of protease inhibitors. Specific labeling was assessed by coincubating unlabeled ET-1, ET-2, ET-3 and other unrelated cytokines. The tissue labeled with 125I-ET-1 was collected by filtration and counted in a gamma counter. For an in vitro autoradiography study, 15 microns cryostat sections were incubated with 125I-ET-1 (0.1 nmol/L). They were fixed, dipped in liquid emulsion and kept for 6 days before development. Membrane counting showed that the binding of 125I-ET-1 to Harder's gland was saturable. Scatchard data analysis revealed one class of binding with a dissociation constant (Kd) of 0.33 nmol/L and a maximal density of binding (Bmax) of 794 attomole/mg of protein. The binding was inhibited most by ET-1, followed by ET-2 and then ET-3 but not by unrelated peptides. Emulsion-dipped slides with sections showed specific high-density labeling mainly over structures identified from serial sections stained by hematoxylin-eosin as the walls of capillaries, arterioles, arteries, and veins of the glands. Less dense binding was found in both white and pink lobes of the gland. No binding was found in fat and connective tissues. The distribution of endothelin action sites in the glandular blood vessels and Harder's gland suggests that the peptide may have a role in the regulation of blood circulation and glandular secretion in the normal rabbit.

Animals↗

Interferon-alpha and interferon-gamma induced modulation of proteins in human corneal fibroblasts.

Little is known about the effects of interferon (IFN) on cell function in the eye. We have analyzed the effect of INF-alpha and IFN-gamma on the expression of proteins in cultured human corneal fibroblasts. Treatment with IFN-alpha increased the synthesis of proteins of 84, 76, 52, and 28 kD and decreased the synthesis of a 72-kD protein. Treatment with IFN-gamma increased the synthesis of proteins of 83, 66, 64, 54, and 47 kD. The effect of IFN-alpha and IFN-gamma were first detected at 5-9 h and 9 h, respectively, after the addition of the IFNs and were maximal at 17 and 24 h, respectively. Most of the changes were seen at doses of 1 x 10(1) to 1 x 10(2) U/ml of IFN-alpha or IFN-gamma and were maximal at 1 x 10(2) to 1 x 10(3) U/ml. Thus, each IFN induced distinct proteins based on apparent molecular weight and isoelectric point. These results show that IFN-alpha and IFN-gamma affect the synthesis of small groups of distinct proteins in human corneal fibroblasts.

Cells, Cultured↗

Conjunctivitis of the newborn.

Infectious conjunctivitis of the newborn is caused by a wide variety of microorganisms. The ocular findings may be part of a widespread systemic infection. Clinical presentations are not diagnostic of the cause, and a microbiologic work-up with cytology, cultures, and microbial sensitivities is mandatory. The selection of specific antimicrobial therapy is based on the findings of laboratory studies. Prophylaxis with silver nitrate solution, 1.0% tetracycline, or 0.05% erythromycin ointment is effective for the prevention of gonococcal and chlamydial conjunctivitis in the newborn.

Anti-Bacterial Agents↗

Indications for penetrating keratoplasty: 1980-1988.

A retrospective analysis was undertaken of the clinical diagnoses of 1594 eyes that underwent penetrating keratoplasty performed in a private-referral corneal practice over a 9-year period, 1980-1988. The seven most common indications for surgery were keratoconus (24.0%), pseudophakic or aphakic bullous keratopathy (21.2%), corneal scarring (13.9%), Fuchs' endothelial dystrophy (12.5%), regraft (8.1%), and herpetic keratitis (5.3%). Keratoconus was the leading indication from 1980 to 1985. From 1985 to 1988, pseudophakic bullous keratopathy became the leading indication and correlates well with known complications associated with closed-loop anterior chamber lenses, which were widely used during the early 1980s. Less frequent indications for penetrating keratoplasty included the following: infectious (nonviral) keratitis (3.5%); acute or chronic ulcerative keratitis (2.7%); interstitial keratitis (1.8%); mechanical trauma (1.5%); other (non-Fuchs') corneal dystrophies (1.4%); congenital opacities (0.8%); and chemical burns (0.5%).

Aphakia↗

Heparin and infarct coronary artery patency after streptokinase in acute myocardial infarction.

Anticoagulant therapy is frequently used after thrombolytic agents in the treatment of acute myocardial infarction (AMI) although it is unclear that such therapy will prevent subsequent infarct vessel reocclusion. The role of duration of heparin therapy in maintaining infarct artery patency was studied retrospectively in 53 consecutive AMI patients who received streptokinase therapy and underwent coronary angiography acutely and at 14 +/- 1 days. Of the 39 patients with initial infarct vessel patency, patency at follow-up angiography was observed in 100% (22 of 22) of those who received greater than or equal to 4 days of intravenous heparin but in only 59% (10 of 17) of those patients who received less than 4 days of heparin (p less than 0.05). Of the 14 patients not initially recanalized after streptokinase, patent infarct-related arteries at follow-up angiography were found in 3 of 8 (38%) treated with greater than or equal to 4 days of heparin therapy but in none of the 6 patients treated for less than 4 days (difference not significant). No significant difference in hemorrhagic complications was noted between the short- and long-term heparin treatment groups. Thus, greater than or equal to 4 days of intravenous heparin therapy after successful streptokinase therapy in AMI is more effective in maintaining short-term infarct vessel patency than a shorter duration of therapy and it may maintain the short-term patency of the infarct vessel in those patients who later spontaneously recanalize.

Angiography↗

Needle growth in Sitka spruce (Picea sitchensis): effects of nutrient deficiency and needle position within shoots.

Needle development and shoot growth were studied in 14- and 20-year-old Sitka spruce (Picea sitchensis (Bong.) Carr.) planted on an oligotrophic peat and fertilized with N, P and K (control), N and P (-K), N and K (-P) or unfertilized (-NPK). Shoot extension, needle number, and the size and number of needle cells were observed throughout the season. Beginning with bud burst in early May, needle growth continued for nine weeks in the control treatment. Needle growth in the mineral deficiency treatments terminated at the same time as in the controls, although it did not begin until up to three weeks later than in the control trees. Needles developed acropetally along the shoot, with basal needles completing their development first. Cell division in needles of control trees lasted about 3 weeks, with cell expansion continuing for a further 3 weeks. Initial mean cell cycle times were shortest for proximal needles following bud burst, but lengthened as the season progressed. Cell number increased from 6600 in the primordia to 200 000 in mature needles. Final needle dimensions and cell number varied according to the position of the needle on the shoot. The largest needles with the most cells were near the middle of the shoot. Relative to the controls, all mineral deficiency treatments reduced shoot length, bud and needle dimensions, and needle cell number, particularly the -P treatment. Potassium and P deficiency treatments reduced primordial cell number by up to 42%, whereas final mean cell size was 30% greater in the -P treatment, 17% greater in the -NPK treatment, but 14% smaller in the -K treatment. Mean cell size was constant for needles at all positions in any particular treatment at any time, so that final needle size was determined by cell number alone. Needle dry weight/fresh weight ratio continued to increase until early August, with significantly higher ratios in the -K and -P treatments than in the controls.

Journal Article↗