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K O'Brien

Publications and source records attributed to K O'Brien.

At least 19 recordsLinked to original sources

Penetrating sledding injuries to the lower torso--2 case reports.

Sledding accidents are frequent and vary in severity. Penetrating sledding injuries are uncommon but may be devastating. Snow-racers--sleds with both steering and braking devices--may be associated with an increased rate of injury. The authors present 2 cases of lower-torso penetrating trauma associated with the use of snow-racers. Both cases involved penetration--of the perineum in one case and the inguinal area in the other--by wooden sticks. Both patients recovered fully after prompt surgical intervention. The authors suggest that the absence of a protective panel at the front of the snow-racer may result in the sledder's lower torso being more exposed to objects encountered while sledding. The injuries reported raise concerns about the safety of modern sleds and the possibility that design changes are needed.

Adolescent

Effect of ex vivo hyperthermia on radiation-induced micronuclei in lymphocytes of cancer patients before and during radiotherapy.

To investigate the effect of ex vivo hyperthermia (HT) and 137Cs-irradiation on micronucleus (MN) production in cytokinesis-blocked lymphocytes, we obtained the peripheral blood samples from the same cancer patients (n=6) before and during fractionated partial-body radiotherapy (xRT). The whole blood cultures were heated at 43.5 degrees C for 60 min, followed by 137Cs irradiation (0-4 Gy). The control cultures from the same patients were incubated at 37 degreesC after being exposed to radiation. The lymphocytes were then stimulated with PHA. Cytochalasin B was applied at 44 h, and lymphocytes were harvested at 72 h. MN frequency was determined on Giemsa-stained slides. We found that in patients before xRT, HT (43.5 degrees C) significantly increased the MN yield (mean+/-SEM) in unirradiated lymphocytes from 15.6+/-2.8 (37 degrees C) to 39.7+/-10.9. Further, in patients either before or during xRT, when the lymphocytes were treated with HT (43.5 degrees C) and combined with ex vivo irradiation, the MN yield (Y) could be estimated by a linear equation Y=C+alphaD. Our findings indicate that as measured by the MN production in cytokinesis-blocked lymphocytes, HT alone at 43.5 degrees C++ induced DNA damage. Moreover, it enhanced the radiation-induced cytogenetic damage. Therefore, the application of HT may impair the T-cell function in cancer patients who are receiving radiotherapy. 1998 Elsevier Science B.V.

Aged

Tracheal intubation without neuromuscular blockade in children: a comparison of propofol combined either with alfentanil or remifentanil.

Forty healthy children, aged between two and 12 years of age undergoing elective surgery where the anaesthetic technique involved tracheal intubation followed by spontaneous ventilation were studied. Induction of anaesthesia was with either alfentanil 15 micrograms.kg-1 or remifentanil 1 microgram.kg-1 followed by propofol 4 mg.kg-1 to which lignocaine 0.2 mg.kg-1 had been added. Intubating conditions were graded on a four point scale for ease of laryngoscopy, vocal cord position, degree of coughing, jaw relaxation and limb movement. All children were successfully intubated at the first attempt. There were no significant differences in the assessments of intubating conditions between the two groups. Arterial blood pressure and heart changes were similar in the two groups with both alfentanil and remifentanil attenuating the haemodynamic response to tracheal intubation. The time taken to resumption of spontaneous ventilation was similar in both groups.

Alfentanil

Sevoflurane compared with halothane for tracheal intubation in children.

We have studied 40 healthy children, aged 3-10 yr, undergoing adenotonsillectomy, in a double-blind, randomized study. Intubating conditions were assessed when the pupils had become small and central after inhalation induction with either 5% halothane and 60% nitrous oxide in oxygen or 8% sevoflurane and 60% nitrous oxide in oxygen. The quality of tracheal intubation was graded according to ease of laryngoscopy, position of the vocal cords, coughing, jaw relaxation and movement of limbs. Fewer children had significant vocal cord movement on laryngoscopy (P < 0.01) and more had ideal intubating conditions when halothane was used (12 of 20 compared with seven of 20; ns). Time to reach the clinical end-point for intubation was reached sooner with halothane (P = 0.015). In all children the trachea was intubated successfully at the first attempt and all remained haemodynamically stable throughout induction.

Adenoidectomy

Induction and emergence in infants less than 60 weeks post-conceptual age: comparison of thiopental, halothane, sevoflurane and desflurane.

We have studied 40 infants with a post-conceptual age of less than 60 weeks undergoing general anaesthesia for herniotomy. Patients were anaesthetized with 1 MAC equivalent values for age and agent and allocated randomly to receive halothane, savoflurane or thiopental for induction, and halothane, sevoflurane or desflurane for maintenance of anaesthesia. At induction, both time to acceptance of a face mask and loss of eyelash reflex were recorded. Emergence times were noted by a blinded observer. Induction and emergence times were similar between the halothane and sevoflurane groups but were consistently shorter in the desflurane group compared with the halothane or sevoflurane groups. There were no problems at extubation or significant apnoea in any group. Induction of anaesthesia in this population was no quicker with sevoflurane than with halothane and the method used for induction did not influence recovery time. Maintenance of anaesthesia with desflurane resulted in a shorter recovery time in infants in whom anaesthesia was induced with halothane or thiopental. Desflurane maintenance may be particularly beneficial in the neonate.

Anesthesia Recovery Period

Cloning, expression, purification, and characterization of the acid alpha-mannosidase from Trypanosoma cruzi.

The acid alpha-mannosidase of Trypanosoma cruzi is a broad-specificity hydrolase involved in the catabolism of glycoconjugates, presumably in the digestive vacuole. We have cloned the alpha-mannosidase gene from a T.cruzi epimastigote genomic library. The alpha-mannosidase gene was determined to be single copy by Southern analysis, and similar sequences were not detected in genomic digests of either Trypanosoma brucei or Leishmania donovani. The coding region was subcloned into the Pichia pastoris expression vector pPICZ, and alpha-mannosidase activity was detected in the medium of induced cultures. The recombinant alpha-mannosidase demonstrated a pH optimum, inhibition by swainsonine, Km, and substrate specificity consistent with the characteristics of the alpha-mannosidase previously purified from T.cruzi epimastigotes. The recombinant enzyme was purified 103-fold from the culture medium of Pichia pastoris and had a native molecular mass of 359 kDa by gel filtration. A combination of SDS-PAGE, deglycosylation with endo H, and NH2-terminal sequencing indicates that the enzyme is originally synthesized as a homodimeric polypeptide that is subsequently cleaved to form a heterotetramer composed of 57 and 46 kDa subunits. A polyclonal antibody raised to the recombinant enzyme was shown to immunoprecipitate the alpha-mannosidase from T.cruzi cell extracts and will be used in future immunolocalization studies.

Amino Acid Sequence

MerMade: an oligodeoxyribonucleotide synthesizer for high throughput oligonucleotide production in dual 96-well plates.

We have designed and constructed a machine that synthesizes two standard 96-well plates of oligonucleotides in a single run using standard phosphoramidite chemistry. The machine is capable of making a combination of standard, degenerate, or modified oligos in a single plate. The run time is typically 17 hr for two plates of 20-mers and a reaction scale of 40 nM. The reaction vessel is a standard polypropylene 96-well plate with a hole drilled in the bottom of each well. The two plates are placed in separate vacuum chucks and mounted on an xy table. Each well in turn is positioned under the appropriate reagent injection line and the reagent is injected by switching a dedicated valve. All aspects of machine operation are controlled by a Macintosh computer, which also guides the user through the startup and shutdown procedures, provides a continuous update on the status of the run, and facilitates a number of service procedures that need to be carried out periodically. Over 25,000 oligos have been synthesized for use in dye terminator sequencing reactions, polymerase chain reactions (PCRs), hybridization, and RT-PCR. Oligos up to 100 bases in length have been made with a coupling efficiency in excess of 99%. These machines, working in conjunction with our oligo prediction code are particularly well suited to application in automated high throughput genomic sequencing.

Chromosome Mapping

Limb defects in homozygous alpha-thalassemia: report of three cases.

Homozygosity for the South-Asian alpha-thalassemia (--SEA/) deletion is a serious hematological condition that results, in most cases, in intrauterine or postnatal death due to anemia and severe hypoxia of prenatal onset. A relationship between congenital abnormalities and intra-uterine hypoxia has been postulated. However, since homozygosity for the (--SEA/) deletion is most common in underdeveloped countries where detailed autopsies are lacking, the incidence of congenital abnormalities among these babies has not been well delineated. We report on three newborn infants, homozygous for the (--SEA/) deletion, who were born with limb defects. We postulate that this combination is the result of prenatal hypoxia which may affect other fetal body organs. This should be taken into consideration when prenatal treatment of affected fetuses, with intrauterine blood transfusion, is suggested.

Adult

Construction and characterization of a one-plasmid system for the controlled expression of genes in mammalian cells by tetracycline.

The two-plasmid system of Gossen and Bujard [Gossen and Bujard (1992) Proc. Natl. Acad. Sci. USA 89, 5547-5551] to express mammalian genes in a tetracycline-repressed fashion was combined into a single-plasmid system. Two variants of this single-plasmid system that differ in the multiple cloning site (MCS) region are described. These vectors were used to stably transfect raf kinase domain into the normal rat kidney epithelial cell line (NRKE) to obtain a conditionally transformed cell line. These vectors were also used to stably transfect wild-type and mutant human p53 into the human osteosarcoma cell line, SAOS-2. Tetracycline repressed gene expression in both cell lines; about 12-fold in NRKE and about 80-fold in SAOS-2 cell line.

Animals

Polycystic ovarian condition in the dehydroepiandrosterone-treated rat model: hyperandrogenism and the resumption of meiosis are major initial events associated with cystogenesis of antral follicles.

BACKGROUND: The purpose of this study was to elucidate the early effects of dehydroepiandrosterone (DHEA) in the polycystic rat model by charting cytological changes in the early antral follicle of the ovary and constructing a serum hormonal profile. Histological examinations of ovaries from DHEA-treated rats for ten consecutive days revealed that the oocyte of antral follicles, ranging from 1.5 mm to 3.4 mm in diameter, had become activated, i.e., had resumed meiosis. Tabulation and statistical analysis revealed a highly significant difference in the percentage of oocyte activation between the ovaries of DHEA-treated and control rats. Granulosa cells associated with those antral follicles included in our statistical analysis showed no evidence of atresia. A few follicles not included in our analysis contained oocytes that had resumed meiosis and whose associated granulosa cells were atretic. The observed resumption of meiosis occurred in the absence of surges of follicle stimulating hormone (FSH) and luteinizing hormone (LH). During meiosis, a period when many oocytes become activated, levels of serum androgens (DHEA, testosterone, and androstenedione) were high, while FSH, LH, and prolactin (PRL) levels did not differ significantly from those in the controls. Follicles that resume meiosis may be members of a group of follicles that produces a signal(s) when the oocyte becomes uncoupled from the granulosa cell. This signal(s) permit(s) a reprogramming of the accompanying granulosa cells of the follicle to engage in certain developmental processes of cystogenesis. Just what cascade of signals is necessary to achieve this selection remains elusive at this time and is the subject of our continuing investigations.

Androstenedione

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