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Biomedical subjects

J E Oliver

Publications and source records attributed to J E Oliver.

At least 19 recordsLinked to original sources

Application of Bruchin B to pea pods results in the up-regulation of CYP93C18, a putative isoflavone synthase gene, and an increase in the level of pisatin, an isoflavone phytoalexin.

Bruchins, mono and bis (3-hydroxypropanoate) esters of long chain alpha,omega-diols, are a recently discovered class of insect elicitors that stimulate cell division and neoplasm formation when applied to pods of peas and certain other legumes. Differential display analysis resulted in the identification of an mRNA whose level was increased by the application of Bruchin B to pea pods. The corresponding amplification product was cloned and sequenced and a full length cDNA sequence was obtained. This cDNA and the gene from which it was derived were assigned the name CYP93C18 based upon sequence similarities to the cytochrome P450 mono-oxygenase CYP93C subfamily, which contains isoflavone synthase genes from legumes. RNA gel blots and quantitative RT-PCR demonstrated that expression of CYP93C18 increased within 8 h of bruchin treatment to a maximum of 100-200-fold of the level in untreated pods, and then declined. The up-regulation of CYP93C18 was followed by an increase in the level of the isoflavone phytoalexin, pisatin. Pisatin was detectable in the bruchin-treated pods after 16 h and reached a maximum between 32 h and 64 h. This, the first report of induction of phytoalexin biosynthesis by an insect elicitor, suggests that Bruchin B not only stimulates neoplasm formation, but also activates other plant defence responses.

Amino Acid Sequence↗

Characterization of a behaviorally active, gender-specific volatile compound from the male asparagus fly Plioreocepta poeciloptera.

Adult male asparagus flies exhibit typical calling behaviors (suggestive of pheromone production) during which they emit a single volatile compound that was identified as isopropyl (S)-5-hydroxyhexanoate. In laboratory bioassays, synthetic samples elicited an arrestant response in females, but did not appear to attract females. On the other hand, the synthetic material attracted conspecific males in olfactometer bioassays.

Age Factors↗

Coordination between donor cell type and cell cycle stage improves nuclear cloning efficiency in cattle.

Several studies have shown that both quiescent and proliferating somatic donor cells can be fully reprogrammed after nuclear transfer (NT) and result in viable offspring. So far, however, no comparative study has conclusively demonstrated the relative importance of donor cell cycle stage on nuclear cloning efficiency. Here, we compare two different types of bovine fetal fibroblasts (BFFs) that were synchronized in G(0), G(1), and different phases within G(1). We show that for non-transgenic (non-TG) fibroblasts, serum starvation into G(0) results in a significantly higher percentage of viable calves at term than synchronization in early G(1) or late G(1). For transgenic fibroblasts, however, cells selected in G(1) show significantly higher development to calves at term and higher post-natal survival to weaning than cells in G(0). This suggests that it may be necessary to coordinate donor cell type and cell cycle stage to maximize overall cloning efficiency.

Animals↗

Cloned cattle derived from a novel zona-free embryo reconstruction system.

As the demand for cloned embryos and offspring increases, the need arises for the development of nuclear transfer procedures that are improved in both efficiency and ease of operation. Here, we describe a novel zona-free cloning method that doubles the throughput in cloned bovine embryo production over current procedures and generates viable offspring with the same efficiency. Elements of the procedure include zona-free enucleation without a holding pipette, automated fusion of 5-10 oocyte-donor cell pairs and microdrop in vitro culture. Using this system, zona-free embryos were reconstructed from five independent primary cell lines and cultured either singularly (single-IVC) or as aggregates of three (triple-IVC). Blastocysts of transferable quality were obtained at similar rates from zona-free single-IVC, triple-IVC, and control zona-intact embryos (33%, 25%, and 29%, respectively). In a direct comparison, there was no significant difference in development to live calves at term between single-IVC, triple-IVC, and zona-intact embryos derived from the same adult fibroblast line (10%, 13%, and 15%, respectively). This zona-free cloning method could be straightforward for users of conventional cloning procedures to adopt and may prove a simple, fast, and efficient alternative for nuclear cloning of other species as well.

Animals↗

Effects of follicular size of cytoplast donor on the efficiency of cloning in cattle.

In cattle, oocytes obtained from follicles smaller than 3 mm in diameter can undergo maturation in vitro, progressing to MII and undergoing fertilization, but are developmentally incompetent. Cytoplasts were prepared from in vitro matured oocytes aspirated from small (1-3 mm) or large (6-12 mm) follicles and fused to serum starved mural granulosa cells. Following activation, reconstructed embryos were cultured for 7 days and classified G1 to G4, before being processed for nuclei counting or transferred to synchronized recipients. Oocytes from small follicles had lower rates of polar body extrusion (59.6 vs. 69%; 731/1230 vs. 608/857) and fusion (71.4 vs. 78.8%; 360/497 vs. 364/465; P < 0.06). There were no differences in total rate of blastocysts development (60 vs. 59.8%; small vs. large), or any grade classification. A significant interaction was detected between follicle size and embryo grade with G3 embryos from small follicles having a greater cell number. Developmental competence of G1 and G2 embryos did not differ at day 27 (48 vs. 46%; 16/33 vs. 17/37; small vs. large). Although there were no differences in fetal size between the two groups, differences in allantois length (53 vs. 86 mm; small vs. large; P < 0.002) and allantois width (9.5 vs. 13 mm; small vs. large; P < 0.06) were seen. No differences in survival to term (2/13 in each group) were observed. These results indicate that cytoplasts from follicles of 1-3 and 6-12 mm in diameter are equally developmentally competent when used in a nuclear transfer procedure.

Animals↗

The efficacy of ergometry determined heart rates for flatwater kayak training.

The aim of this study was to investigate the use of incremental ergometry determined heart rate training intensities for the control of kayak ergometer and open water kayak training. Eight well-trained male kayakers completed a maximal incremental exercise test on an air-braked kayak ergometer for the determination of LT(1) (the power output at which blood lactate concentration increased by > or = 1 mmol x L(-1)), the associated heart rate (HR-LT(1)), VO(2)peak, maximal heart rate and maximal aerobic power. Subjects then performed 20 min trials of kayak ergometry (E), open water kayaking in a single kayak (K1) and open water kayaking in a four-seat kayak (K4) at HR-LT(1). During the three trials, heart rate was continuously measured, and blood lactate concentration, rating of perceived exertion (RPE) and stroke rate were determined every 5 min. In all trials, exercise at HR-LT(1) resulted in stable blood lactate concentrations and a stable RPE. Comparison of the three trials demonstrated that the only difference was for RPE, which was lower in (K4) than in (E), (p < 0.05). The results demonstrate that the prescription of HR-LT(1) elicits similar blood lactate concentrations during kayak ergometer and open water kayak training in both single and team boats.

Adult↗

The long-term outcome of glaucoma filtration surgery.

PURPOSE: To determine the long-term outcome of glaucoma filtration surgery in preserving vision. Visual loss from progressive glaucomatous damage and from complications of surgery, both short and long term, were included. METHODS: A retrospective, community-based, longitudinal study of residents of Olmsted County, Minnesota, who were newly diagnosed with open-angle glaucoma between 1965 and 1980 and underwent filtration surgery in these or subsequent years through 1998. Intraocular pressure (IOP), visual acuity, visual fields, and progression to legal blindness were monitored. Kaplan-Meier analysis was used to determine the cumulative probabilities of changes in these parameters. RESULTS: 73 eyes of 49 patients underwent conventional filtration surgery. Analysis of the first eye having surgery revealed a mean preoperative IOP of 27.6 +/- 8.5 mm dropping to 16.7 +/- 5.6 mm at year one, and remaining in this range throughout follow-up (14.7 +/- 3.0 mm at 10 years; with or without use of medications). The probability of progression to blindness was 46% at 10 years after surgery, as calculated by Kaplan-Meier analysis. Eyes going blind had a postoperative IOP equal to or lower than those not becoming blind (14.0 +/- 4.4 vs. 15.4 +/- 3.0 at postoperative year 10). Eyes going blind had more advanced field loss at the time of surgery, with scotomas above and below the horizontal axis, than eyes not going blind, which had scotomas in only one hemifield. Three patients developed late bleb leaks; two patients developed endophthalmitis. The probability of undergoing cataract surgery was 37% by 10 years postoperatively, which did not differ significantly from the cohort of patients not undergoing surgery at a comparable time point. CONCLUSIONS: Filtration surgery was associated with a 54% probability of preservation of vision from progression to legal blindness at 10 years after surgery. Patients becoming blind had more advanced field loss at the time of surgery; IOP was similar between those going blind and those retaining vision.

Aged↗

Results of cystometry and urethral pressure profilometry in dogs sedated with medetomidine or xylazine.

OBJECTIVE: To compare effects of medetomidine and xylazine hydrochloride on results of cystometry and micturition reflexes in healthy dogs and results of urethral pressure profilometry (UPP) in sedated and conscious dogs. ANIMALS: 20 dogs. PROCEDURES: Urodynamic testing was performed 6 times in each dog (3 times after administration of xylazine [1 mg/kg of body weight, IV] and 3 times after administration of medetomidine (30 microg/kg, IM). Before each episode of sedation, UPP was performed. Heart and respiratory rates and indirect blood pressures were recorded prior to and 5, 10, 20, and 30 minutes after injection of sedative. Cystometry measurements included threshold volume, threshold pressure, and tonus limb. The UPP measurements included maximal urethral closure pressure (MUCP), functional profile length, and, in male dogs, plateau pressure. RESULTS: Mean MUCP was decreased markedly in xylazine- and medetomidine-sedated dogs. Xylazine and medetomidine also decreased plateau pressure in male dogs. The MUCP measurements were consistent among days for conscious and xylazine-sedated dogs but were inconsistent for medetomidine-sedated female dogs. The proportion of valid cystometry measurements was greater for xylazine (39 of 60) than for medetomidine (27 of 60). Cystometry was considered invalid when bladder pressure reached 30 cm H2O without initiation of a micturition reflex. CONCLUSIONS AND CLINICAL RELEVANCE: Medetomidine and xylazine have similar effects on measurement of UPP and cystometry. Medetomidine was less consistent among days for UPP in female dogs and produced fewer valid cystometry tests, compared with xylazine. For urodynamic evaluations, medetomidine administered IM cannot be substituted for xylazine administered IV.

Animals↗

Bruchins: insect-derived plant regulators that stimulate neoplasm formation.

Pea weevil (Bruchus pisorum L.) oviposition on pods of specific genetic lines of pea (Pisum sativum L.) stimulates cell division at the sites of egg attachment. As a result, tumor-like growths of undifferentiated cells (neoplasms) develop beneath the egg. These neoplasms impede larval entry into the pod. This unique form of induced resistance is conditioned by the Np allele and mediated by a recently discovered class of natural products that we have identified from both cowpea weevil (Callosobruchus maculatus F.) and pea weevil. These compounds, which we refer to as "bruchins," are long-chain alpha,omega-diols, esterified at one or both oxygens with 3-hydroxypropanoic acid. Bruchins are potent plant regulators, with application of as little as 1 fmol (0.5 pg) causing neoplastic growth on pods of all of the pea lines tested. The bruchins are, to our knowledge, the first natural products discovered with the ability to induce neoplasm formation when applied to intact plants.

Animals↗

Lipid composition of the extracellular matrix of Botrytis cinerea germlings.

Six simple lipid classes (mono-, di- and tri-acylglycerols, free fatty acids, free fatty alcohols and wax esters) were identified by TLC in the extracellular matrix of Botrytis cinerea germlings and the molecular components of each class were characterized using GC-MS. The relative amounts of fatty acids and fatty alcohols within each lipid class were determined by GC-FID. Over all the lipid classes, the most abundant saturated fatty acids were palmitic (ca. 30%) and stearic acid (ca. 22%). Palmitoleic and oleic acids made up ca. 21% and 24% (respectively) of the free fatty acids, while erucic (ca. 4.1%) and linoleic (ca. 3.6%) acids were the most abundant unsaturated fatty acids in the acylglycerides. The acylglycerides also contained almost 35% long chain fatty acids (C20:0 to C28:0). Six fatty acids were identified which had odd-numbered carbon chain lengths (C15:0, C17:0, C19:0, C21:0, C23:0 and C25:0). Of these, pentacosanoic acid made up almost 14% of the fatty acids in the acylglycerides. Three methyl-branched chain fatty acids, namely isopalmitic, isoheptadecanoic and anteisopalmitic, were identified in the ECM, all in small amounts. Of the fatty alcohols identified, only palmityl and stearyl alcohols were found in the free form (ca. 57% and 43%, respectively) but arachidyl alcohol (ca. 47%) and 1-octacosanol (ca. 30%) were the most abundant fatty alcohols found in the wax ester fraction.

Botrytis↗

Studies on the measurement of phospholipase A2 (PLA2) and PLA2 inhibitor activities in ram semen.

Extraction with Tris-citrate or Tris-NaCl-EGTA improved the yield of phospholipase A2 (PLA2) from ram semen by 40-50 fold over the previously recommended method of extraction by dilute (0.18 N) sulphuric acid. The enzyme activity in the citrate extract deteriorated more rapidly than in Tris-NaCl-EGTA. The semen PLA2 activity was optimum at pH 8.0, heat sensitive at 70 degrees C for 30 min, activated by Ca2+ (although approximately 60% activity was also found in the absence of calcium) and did not exist as a pro-enzyme. The semen PLA2 activity was equally distributed among the sperm and seminal plasma (SP) components of ram semen. However, the low levels of PLA2 activity in the SP of vasectomised rams tend to suggest that PLA2 in the SP fraction may have originated from testicular or epididymal secretions or leakage, from sperm. PLA, in sperm exists as a large molecular weight aggregate, whereas in SP it is present as a smaller aggregate. In addition to PLA2, semen also contained PLA2 inhibitor activities. Inhibition was observed against PLA2s from bee venom, pig pancreas and oviductal extracts. The inhibitory activity is presumed to be due to a large molecular weight protein as the inhibitor activity was not extracted in a chloroform:methanol (2:1; v/v) mixture, it was non-dialysable, precipitated by 10% trichloroacetic acid and destroyed by proteases. The inhibitor activity was distributed in various molecular weight fractions of sperm, SP and SP from vasectomised rams.

Animals↗

Stress leak point pressures and urethral pressure profile tests in clinically normal female dogs.

OBJECTIVE: To develop a stress leak point pressure (LPP) test for dogs, determine LPP for continent female dogs, and determine urethral pressure profile (UPP) values for nonanesthetized, continent female dogs. ANIMALS: 22 continent female dogs weighing from 21 to 29 kg. PROCEDURE: A standard UPP test and a modification of the LPP test used in women were performed on all dogs. On 3 occasions, dogs underwent UPP testing while awake. They then were anesthetized with propofol, and LPP was measured at bladder volumes of 75, 100, and 150 ml. For LPP tests, abdominal pressure was applied by inflating a human blood pressure cuff placed around the dog's abdomen. LPP were recorded through a urethral catheter (bladder LPP) and a rectal balloon catheter (abdominal LPP). RESULTS: Mean +/- SD and median maximal urethral closure pressure was 110.1+/-20.2 and 109.0 cm water, respectively. Mean bladder LPP for the 75, 100, and 150 ml bladder volumes was 172.4 cm water. Significant differences among LPP for the 3 bladder volumes were not detected. CONCLUSIONS: Stress LPP can be recorded in female dogs.

Animals↗

Vision-specific function and quality of life after cataract extraction in south India.

PURPOSE: To assess visual and overall patient function after intracapsular (ICCE) and extracapsular (ECCE) cataract extraction in rural South India. SETTING: Aravind Eye Hospital, Tirunelveli, Tamil Nadu, South India, and the Dana Center for Preventive Ophthalmology, Wilmer Eye Institute, Baltimore, Maryland, USA. METHODS: This study evaluated preoperative visual acuity and demographic information and postoperative visual acuity and functional status measures in 71 patients having ECCE with posterior chamber intraocular lens (IOL) placement and 73 patients having ICCE with aphakic spectacle correction at Aravind-Tirunelveli Eye Hospital, Tamil Nadu, India. The principal outcomes assessed were visual acuity; quality-of-life score (possible range 0 to 100%); visual function measurement (possible range 0 to 100%). RESULTS: Patients in the ECCE group scored 10.17 (P = .0001) points higher than those in the ICCE group on the visual function scale after adjustment for differences in age, sex, level of education, marital status, residence, and type of employment. The ECCE group scored 7.69 points higher on visual function when adjusting for the differences in best corrected visual acuity, which was also better in the ECCE group. In the quality-of-life assessment, 77.1% in the ECCE group and 46.6% in the ICCE group scored 90% or better (OR 3.85; P = .006). CONCLUSIONS: Patients in rural south India having ECCE with posterior chamber IOL implantation obtained better postoperative visual function, quality of life, and visual acuity than those receiving ICCE with aphakic spectacle correction. These differences, which were not significantly affected by adjustment for age, sex, education, marital status, type of residence, and occupation, indicate that ECCE is clearly superior to ICCE.

Adult↗

Use of partial prostatectomy for treatment of prostatic abscesses and cysts in dogs.

OBJECTIVE: To determine whether dogs had prostatic disease, urinary incontinence, or urinary tract infection 1 year after partial prostatectomy to treat prostatic abscesses and cysts. DESIGN: Prospective study. ANIMALS: 20 male dogs with prostatic abscesses or cysts. Fifteen dogs had evidence of urinary tract infection. Only 8 dogs urinated normally; the remainder dribbled, had obstructions, or required medical treatment. PROCEDURE: Partial prostatectomy was performed on each dog. Sexually intact dogs (n = 12) also were castrated. RESULTS: None of the dogs had return of prostatic cystic enlargement or clinical signs of prostatic disease during the first year after surgery. Two dogs were euthanatized within 1 year after surgery, with 1 dog having prostatic enlargement and adenocarcinoma and 1 dog having unrelated lymphosarcoma. Fifteen dogs were continent. The remaining 5 dogs urinated normally but had intermittent and minor incontinence. Eleven dogs had no signs of infection 1 year after surgery, 5 had pyuria or positive urine bacteriologic culture results, 2 did not have urinalysis performed, and 2 were euthanatized. CLINICAL IMPLICATIONS: Dogs with severe prostatic abscesses or cysts and infections can be successfully treated by partial prostatectomy with an ultrasonic surgical aspirator and castration, resulting in long-term disease resolution. Although most dogs with severe prostatic disease do not urinate normally before surgery, nearly all dogs resume normal micturition after partial prostatectomy. Postoperative results of partial prostatectomy appear to be better than those of previous drainage techniques for treatment of prostatic cavitary disease.

Abscess↗

Motility of ram spermatozoa during storage in a chemically-defined diluent containing antioxidants.

The effects of five antioxidants--Vitamin E (VE), butylated hydroxyanisole (BHA), n-propyl gallate (n-PG), deferoxamine mesylate (Desferal) and catalase (EC 1 . 11 . 1 . 6)--on the maintenance of motility of ram spermatozoa in a chemically-defined ram semen diluent (RSD-1) have been evaluated. VE, n-PG and Desferal inhibited spermatozoal motility. The relative inhibition (i.e., ratio of change in % motility over 24 h between the treatment group and the corresponding control) at equimolar concentrations (100 microM) of Desferal, VE and n-PG were 1.6, 1.8 and 3.6 respectively. BHA had no effect at 10 microM but at lower concentrations, gave a slight improvement in motility in freshly diluted spermatozoal samples and in those stored for 1 day at 15 degrees C. The addition of catalase to RSD-1 was also ineffective in improving the motility of spermatozoa. The lack of beneficial effects of the tested antioxidants suggests that RSD-1 itself may destroy reactive oxygen species and the antioxidant activity of RSD-1 components requires further study.

Animals↗

Clinical, hematologic, and biochemical features of a syndrome in Bernese mountain dogs characterized by hepatocerebellar degeneration.

Seven related Bernese Mountain Dogs developed a syndrome Characterized by progressive cerebellar and hepatic disease. Clinically, stiffness in the hind limbs, mild incoordination, and a slight head tremor were first noticeable when pups were 4 to 6 weeks old. The condition progressed, causing pups to assume a wide-based stance. Other signs included head bobbing, spontaneous nystagmus, and, finally, paresis. Hematologic findings included leukocytosis with a left shift; normocytic, normochromic anemia; hypoproteinemia, low serum creatinine, and urea nitrogen concentrations; excessive fasting plasma ammonia concentration; and an increase in concentration of serum bile acids. Portal venography performed on 1 dog revealed a small liver and extensive extrahepatic varicosities. Necropsy revealed cerebellar hypoplasia, nodular liver, extensive abdominal varicosities, and ascites. Histologically, degeneration and depletion of Purkinje's cells and vacuolation, degeneration, and nodular regeneration of hepatic tissues were evident. Preliminary analysis of the pedigree was suggestive of an autosomal recessive pattern of inheritance.

Animals↗