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Biomedical subjects

A King

Publications and source records attributed to A King.

At least 217 records · Page 12Linked to original sources

European multicentre evaluation of a commercial system for identification of methicillin-resistant Staphylococcus aureus.

A commercial system for the rapid detection of methicillin-resistant Staphylococcus aureus, the BBL Crystal MRSA test (C-MRSA ID; Becton Dickinson, USA), was evaluated prospectively and compared with a polymerase chain reaction test for the presence of the mecA gene. Ten European centres tested a total of 676 isolates of Staphylococcus aureus from blood cultures. The system correctly identified 661 (97.8%) isolates within 4 h. All but three mecA gene-negative isolates (99.4% specificity) yielded a negative C-MRSA ID reaction, and 158 of 170 mecA gene-positive isolates were accurately detected (92.9% sensitivity). After repeated testing of discrepant results, sensitivity and specificity increased to 99% and 100%, respectively.

Bacteriological Techniques↗

Human decidual leukocytes do not proliferate in response to either extravillous trophoblast or allogeneic peripheral blood lymphocytes.

The maternal leukocytes present in human decidua during the early weeks of pregnancy are composed of CD56+ NK cells (approximately 80%), CD3+ T cells (approximately 10%), and CD14+ macrophages (approximately 10%). These leukocytes are present in large numbers at the implantation site where trophoblast invades. The migrating extravillous trophoblast (EVT) cells express HLA class I antigens HLA-G and HLA-C. To examine whether EVT or other allogeneic cells are capable of inducing a local proliferative response in maternal decidual leukocytes, we have assayed [3H]thymidine incorporation by decidual leukocytes co-cultured with irradiated trophoblast or peripheral blood lymphocytes (PBL). No proliferative response was seen in decidual leukocytes either to trophoblast or to PBL as stimulators. In addition trophoblast did not stimulate a proliferative response in PBL. These findings indicate that extravillous trophoblast is not capable of initiating a classical immune response at the implantation site.

Cell Division↗

Immune phenotype and cytotoxic activity of lymphocytes from human term decidua against trophoblast.

Flow cytometric data were used to compare the phenotype of term decidual lymphocytes and peripheral blood lymphocytes. Unlike peripheral blood lymphocytes, a significant percentage of CD3+, CD4+, CD8+ and CD16+ term decidual lymphocyte populations expressed the CD69 activation marker. The relative proportions of CD38 in CD3+, CD4+ and CD8+ populations were more than twice as large in term decidual lymphocytes as in peripheral blood lymphocytes. As reported for early decidual lymphocytes, the expression of CD38 and CD69 by term decidual lymphocytes suggests that these cells are also regionally activated. However, term decidual lymphocytes showed no spontaneous cytotoxicity against normal trophoblast or its tumoral counterpart, JEG cells. After stimulation with interleukin-2, these lymphocytes became cytotoxic, as did peripheral blood lymphocytes. The relevance of this latter result to the immune control of the physiological and pathological invasion of the decidua by the trophoblast is discussed.

ADP-ribosyl Cyclase↗

Catheter-induced atrioventricular nodal block during radiofrequency ablation.

This study examined the incidence and significance of catheter-induced atrioventricular nodal block (AVNB) during a radiofrequency ablation procedure that uses stiff large-tip steerable ablation catheters. AVNB was noted in 10 (1.6%) of 613 consecutive patients undergoing radiofrequency ablation therapy for atrioventricular nodal (AVN) reentrant tachycardia (592 patients) or atrioventricular reentry tachycardia incorporating a midseptal accessory pathway (21 patients). Of these 10 patients, 9 underwent AVN modification for AVN reentrant tachycardia and 1 for ablation of a midseptal accessory pathway. One patient had two episodes of AVNB during two sessions undertaken because of recurrence of tachycardia. No patient had a preexisting conduction defect before the study. In all 10 patients, AVNB was transient, and it lasted for a mean of 9.1 +/- 19 minutes. It occurred during positioning of the ablation catheter in the junctional area before (8 patients) or after (2 patients) the start of radiofrequency current applications. Complete AVNB was noted on six occasions, second-degree AVNB on four occasions, and first-degree AVNB on one occasion. All blocks were associated with narrow QRS ventricular beats and with a site of block proximal to the His bundle. The mean ventricular heart rate during AVNB was 60 +/- 23 beats/min. Two patients had transient asystole, with one having loss of consciousness. No patient required special treatment for heart block. One-to-one conduction resumed after repositioning of the catheters, and the subsequent ablation procedure was successfully completed in 8 of the 10 patients. During a follow-up of 20 +/- 12 months, none of the patients had severe dizziness or syncope, and none required implantation of a permanent pacemaker. In conclusion, transient AVNB due to mechanical injury occurs during positioning of a stiff large-tip steerable ablation catheter in the junctional area. Delivery of radiofrequency current to the site that provokes catheter-induced AVNB should be avoided.

Adolescent↗

Radiofrequency ablation therapy of the posteroseptal accessory pathway.

Among 652 patients with Wolff-Parkinson-White syndrome who underwent radiofrequency ablation in this laboratory, 139 (21%) were found to have a total of 146 posteroseptal accessory pathways. Ablation was conducted by the regular transvenous or transaortic approach; ablation from cardiac venous structures was used only if regular approaches were unsuccessful. Of the 146 posteroseptal accessory pathways, 94 were successfully ablated from the left posteroseptal region and 45 from the right posteroseptal region. In 3, successful ablation of the accessory pathway required delivery of the current to the proximal coronary sinus, and in 1 it required delivery of the current to both the atrial and ventricular aspects of the tricuspid valve at the right posteroseptum. Thus, the accessory pathway was successfully ablated in 143 (98%) of 146 instances or in 136 (98%) patients. In 3 patients, ablation was unsuccessful despite delivery of current to the left posteroseptum, the right posteroseptum, the proximal coronary sinus, and the middle cardiac vein. Seventy-seven (57%) patients with an initial success, including 9 patients with resumed preexcitation or recurrence of paipitations, underwent a follow-up electro-physiologic study 90 +/- 72 days after ablation. Of these 9 patients, the initial successful ablation site was the right posteroseptum in 7 and the left posteroseptum in 2. The accessory pathways were ablated successfully by subsequent trials in 8 patients, whereas in 1 the accessory pathway was severely damaged. Thus radiofrequency ablation of posteroseptal accessory pathways can be achieved by the regular transvenous or transaortic approach; delivery of current to the coronary sinus or middle cardiac vein is unnecessary in most patients.

Adult↗

Polymerase chain reaction enzyme-linked immunosorbent assay detection of mycoplasma consensus gene in sperm with low oocyte penetration capacity.

OBJECTIVE: To determine the prevalence of mycoplasmas in washed sperm and to compare the penetration of zona-free hamster oocytes by sperm with and without mycoplasmas. DESIGN: Prospective comparative study. SETTING: Clinical and academic research environment. PATIENT(S): Semen from 34 male patients. INTERVENTION(S): Specimens were divided, Percoll washed, and scanned for differences in kinematic parameters. Sperm DNA was extracted and assayed for mycoplasma DNA using the polymerase chain reaction-ELISA method targeting the consensus gene of 15 mycoplasma species. Remaining sperm were processed by centrifuge, Percoll, or TEST (TES and Tris) Yolk Buffer (TYB) and assessed for penetration capacity. MAIN OUTCOME MEASURE(S): Detection of mycoplasma DNA. RESULT(S): Mycoplasma DNA was detected in 29.4% of the Percoll-washed sperm. The penetration of oocytes by mycoplasma-positive sperm (59.5% +/- 17.3%; mean +/- SEM) was less than mycoplasma-negative sperm (86.8% +/- 5.4%) in the TYB-processed group. CONCLUSION(S): Mycoplasma DNA is demonstrated in almost a third of the Percoll-washed sperm. Because there were no other cell types except sperm, the results suggest that the mycoplasmas were either internalized or attached to the membranes. The reduced penetration by mycoplasma-positive sperm after 48-hour TYB suggest mycoplasmas required time to affect sperm function. Similarities between hypo-osmotic swelling and between kinematic parameters suggest that the mechanism does not involve differences in membrane integrity and in motility patterns.

Animals↗

High-level linguistic disturbances subsequent to childhood closed head injury.

High-level language functioning was assessed in a group of 11 children with severe closed head injury (CHI). Performance was examined on measures of figurative language, oral expression, inferencing, and interpretation of ambiguous sentences, and was compared with that of a control group matched for age, gender, and handedness. The children with CHI demonstrated inferior performance on the oral expression, ambiguous sentences, and figurative language tests; however, performance on the inferencing task appeared intact. These findings are discussed with respect to the underlying nature of the observed impairments. Both linguistic and cognitive factors are considered. It is concluded that a complex interaction of cognitive and linguistic impairments experienced by children with CHI may underlie the performance deficits identified in the current study.

Child↗

Immunology of human implantation: an evolutionary perspective.

The concept of the fetus as an allograft, which has been much cherished by reproductive immunologists, is in need of re-appraisal. We speculate that the relationship between the allogeneic conceptus and mother is not governed by the laws of classical transplantation immunity. Instead, this relationship is more akin to the allorecognition system seen in invertebrates.

Animals↗

Analysis of the flow cytometer stain Hoechst 33342 on human spermatozoa.

Several procedures exist for processing sperm cells for sex preselection. Flow cytometric separation using the fluorochrome stain Hoechst 33342, chemically known as bisbenzimide, is the most promising. The objective of this study was to determine the effect of bisbenzimide on spermatozoa assessed by means of the sperm survival test and to analyse the beta-globin gene in sperm DNA after exposure to increasing concentrations of bisbenzimide. Donor (n = 16) sperm specimens were pooled and washed in a discontinuous Percoll gradient 95:47%, divided and incubated in tubes containing bisbenzimide at concentrations 0 (control), 0.9, 9, 90, 900 and 9000 microM at 25 degrees C and scanned on a computer-aided sperm motility analyser at 0, 1, 4 and 24 h. Spermatozoa were also incubated in a known mutagen, ethidium bromide, as positive control. After 24 h of incubation, the treated sperm cells were processed through DNA extraction and polymerase chain reaction (PCR) performed with primers targeting the beta-globin gene. The amplified DNA products were analysed for evidence of mutation in 5% polyacrylamide gel electrophoresis and 20:80 denaturing gradient gel electrophoresis (DGGE) and further confirmed in 30:40 DGGE. The results showed complete cessation of motility in sperm incubated in the presence of 900 microM or higher concentrations of bisbenzimide. The beat cross frequency sperm parameter was significantly different at the 90 microM or higher concentration of bisbenzimide compared with the control. At concentrations < 900 microM bisbenzimide, there were no differences in the remaining sperm kinematic parameters (percentage rapid progressive, percentage total progressive, sperm velocities, linearity, straightness, amplitude of lateral head displacement and percentage hyperactive motility). PCR and DGGE analyses of spermatozoa treated with bisbenzimide showed no evidence of mutation in the representative region of the beta-globin gene at concentrations < 900 microM. The data suggest an inhibitory effect of bisbenzimide on human sperm motility at 900 microM or higher concentrations of bisbenzimide. The decrease in sperm motility and rapid progression were not due to changes in pH. Point mutation in the representative region of the beta-globin gene in human spermatozoa was detected only at high concentrations (> or = 900 microM) of bisbenzimide. The data suggest that incubating sperm in low concentrations of bisbenzimide (< 90 microM) for up to 24 h does not significantly affect all the sperm kinematic parameters including the beat cross frequency parameter when compared with the control.

Benzimidazoles↗

Evaluation of oestrogen and progesterone receptor expression in uterine mucosal lymphocytes.

Expression of the oestrogen and progesterone receptors on uterine mucosal leukocytes has been examined by dual immunohistology. Neither the oestrogen receptor nor the progesterone receptor was expressed by lymphocytes, macrophages or the distinctive population of uterine natural killer (NK) cells. Although the accumulation and survival of these NK cells appears to be hormonally dependent, the effects must therefore be indirect.

Decidua↗

Incidence of gastroesophageal reflux with whey- and casein-based formulas in infants and in children with severe neurological impairment.

Ten exclusively gastrostomy-fed, neurologically impaired children (4.5-14.5 years old) with gastroesophageal reflux were randomly assigned to receive feedings with either a casein- or a whey-based formula for 48 h each and then crossed over to the other formula. One 24-h pH probe study each was performed while being fed casein- and whey-based formula, respectively. There was a significant reduction in episodes and duration of gastroesophageal reflux while consuming the whey-based formula (p < 0.05). Whey-based feedings should be considered an additional tool in conjunction with other antireflux measures to treat gastroesophageal reflux more effectively in children with severe neurological impairment. A similar study was also conducted involving 14 infants (3-12 months old) with documented gastroesophageal reflux using 24-h pH probe monitoring while consuming a casein-based formula. The formula was changed to a whey-based formula and the pH probe study repeated within 3-5 days. Four infants showed improvement and the rest showed either deterioration (1/14) or comparable results (9/14). The reduction in the mean number of episodes or duration of gastroesophageal reflux with the whey-formula was not significantly different from that with the casein-based formula (p > 0.05). Based on these findings, generalized recommendations for the use of whey-based formula in infants with gastroesophageal reflux cannot be made.

Adolescent↗

Functions of human decidual NK cells.

The main population of lymphocytes found in the human decidua during early pregnancy are NK-like cells with a distinctive phenotype, CD56bright CD16- CD3-. These cells are in close association with invading trophoblast that may be their in vivo target. We have examined three aspects of decidual NK function in vitro: cytotoxicity, proliferation, and cytokine production. The functional assays indicate uterine lymphocytes differ fundamentally from both PBL and even from classical circulating NK cells. Their role in the establishment of normal pregnancy remains unknown.

Cytotoxicity, Immunologic↗

Thrombin activates the sarcolemmal Na(+)-H+ exchanger. Evidence for a receptor-mediated mechanism involving protein kinase C.

Thrombin can activate the plasma membrane Na(+)-H+ exchanger in a variety of noncardiac cells. We have studied (1) the effect of thrombin on the activity of the sarcolemmal Na(+)-H+ exchanger in freshly isolated quiescent ventricular myocytes from the adult rat heart and (2) the signaling mechanism(s) underlying any effect. Reverse-transcription polymerase chain reaction analysis revealed thrombin receptor mRNA expression in a myocyte-enriched cell preparation. As an index of Na(+)-H+ exchanger activity, acid efflux rates (JHS) were determined in single myocytes (n = 4 to 11 per group) loaded with the pH-sensitive fluoroprobe carboxy-seminaphthorhodafluor-1 after two consecutive intracellular acid pulses (induced by transient exposure to 20 mmol/L NH4Cl) in bicarbonate-free medium. At a pHi of 6.9, JH did not change significantly during the second pulse relative to the first in control cells. However, when the second pulse occurred in the presence of 0.2, 1, or 5 U/mL thrombin, JH increased by 30%, 62% (P < .05), and 87% (P < .05), respectively. A hexameric thrombin receptor-activating peptide (SFLLRN) mimicked the effect of thrombin and increased JH by 73% (P < .05) at 25 mumol/L. In contrast, an inactive control peptide (FLLRN) was without effect at 25 mumol/L. In cells pretreated with 100 nmol/L GF109203X or 5 mumol/L chelerythrine (protein kinase C inhibitors), neither 5 U/mL thrombin nor 25 mumol/L SFLLRN produced a significant increase in JH. In the presence of 10 mumol/L HOE-694 (a Na(+)-H+ exchanger inhibitor), pHi did not recover after an acid load, even during exposure to 5 U/mL thrombin or 25 mumol/L SFLLRN, confirming that the Na(+)-H+ exchanger was the primary acid efflux mechanism under the conditions used. Neither 5 U/mL thrombin nor 25 mumol/L SFLLRN affected resting pHi and Ca2+ or background acid loading. We conclude that (1) adult rat ventricular myocytes express a functional thrombin receptor, whose stimulation results in increased activity of the sarcolemmal Na(+)-H+ exchanger, and (2) this effect appears to occur through a protein kinase C-mediated mechanism.

Animals↗