Biomedical subjects

D J Brown

Publications and source records attributed to D J Brown.

At least 109 records · Page 6Linked to original sources

PubMed1995

T cell activation by Theileria annulata-infected macrophages correlates with cytokine production.

A major feature of the pathology induced by Theileria annulata is acute lymphocytic proliferation, and this study investigates the mechanisms underlying the intrinsic ability of T. annulata-infected monocytes to induce naive autologous T cells to proliferate. Different T. annulata-infected clones expressed different but constant levels of MHC class II, varying from < 1.0 x 10(5) to 1.5 x 10(6) molecules/cell, as measured by saturation binding. However, no correlation was found between the level of MHC class II expression and levels of induced T cell proliferation. Theileria annulata-infected cell lines and clones were assayed for cytokine mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR). The infected cells assayed produced mRNA specific for IL-1 alpha, IL-1 beta, IL-6, IL-10 and tumour necrosis factor-alpha (TNF-alpha), but not IL-2 or IL-4. One clone (clone G) did not produce mRNA for TNF-alpha. The degree of T cell proliferation induced by infected cells was directly correlated with the amount of mRNA produced for the T cell stimulatory cytokines IL-1 alpha and IL-6, as assessed by a semiquantitative technique. In contrast, cells infected with the related parasite T. parva produced mRNA for IL-1 alpha, IL-2, IL-4, IL-10 and interferon-gamma (IFN-gamma). Since T. parva-infected cells also induce naive autologous T cell proliferation, it seems likely that the production of IL-1 alpha by cells infected with either parasite is a major signal for the induction of non-specific T cell proliferation.

Animals↗

PubMed1994

Opinions of general practitioners in Nottinghamshire about provision of intrapartum care.

OBJECTIVE: To examine the beliefs of general practitioners concerning intrapartum care. DESIGN: Postal questionnaire survey. SUBJECTS: All general practitioners with patients in Nottinghamshire Family Health Services Authority in September 1993. MAIN OUTCOME MEASURES: General practitioners' current involvement in maternity care, and beliefs on intrapartum care. RESULTS: Of 694 general practitioners sent questionnaires, 550 (79.2%) replied. 529 of these were on the obstetric list; 437 had not attended a delivery in the past 12 months; 36 had attended two or more; 358 general practitioners did not wish to provide more intrapartum care; 349 did not feel competent to do so. Reasons for not wanting to provide intrapartum care included current workload (453), disruption to personal life (407), and the fear of litigation (377). General practitioners who already booked women for home delivery were more likely to wish to do more deliveries (62/42 v 61/316, chi 2 = 85.3; P < 0.0001) and to have more positive attitudes towards increasing women's choice in maternity care (90/22 v 195/151, chi 2 = 227; P < 0.0001). CONCLUSIONS: The involvement of general practitioners in intrapartum care in Nottinghamshire is low, and most general practitioners are unwilling to increase their role. However, general practitioners who already book for home delivery are keen to do more.

Attitude of Health Personnel↗

PubMed1994

Collection of semen from men in acute phase of spinal cord injury.

In chronic spinal cord injury, semen obtained by assisted ejaculation is usually abnormal. We have assessed electroejaculation early after injury in seven patients. There were no adverse effects. Initial samples contained few or no spermatozoa but as patients emerged from spinal shock, semen improved and five had specimens cryopreserved. Thereafter sperm motility and viability decreased towards the pattern of chronic spinal cord injury by day 16. Cryopreservation was not possible in one patient with many medical complications and another who started electroejaculation 15 days after injury. Semen storage within the first 2 weeks after spinal cord injury is recommended for future fertility treatment.

Acute Disease↗

PubMed1994

Remodelling of a displaced phalangeal neck fracture.

Phalangeal neck fractures are uncommon in children. When these injuries to the proximal and middle phalanges are displaced and not treated operatively the fracture may heal in a malunited position with loss of motion at the IP joint. Remodelling in the area of the phalangeal neck is thought to be reduced because of its distance from the physis. In cases of malunion osteotomy of the phalangeal neck may be required to restore anatomy and motion. A case is described which demonstrates complete remodelling of a displaced middle phalangeal neck fracture in a child and recovery of a normal range of motion without operative intervention.

Bone Remodeling↗

PubMed1994

A simple technique to prevent retrograde ejaculation during assisted ejaculation.

The aim of this study was to develop a technique which would prevent retrograde ejaculation in chronic spinal cord injured (SCI) patients undergoing vibration and electroejaculation procedures. A balloon catheter was used to tamponade the bladder neck in 12 patients who underwent 100 assisted ejaculation procedures. Antegrade ejaculations were collected on all occasions with no incidences of urine contamination and no sperm were seen in post ejaculatory urine. Silicone catheters had minimal effects on sperm motility and viability. All lubricant gels were found to adversely affect sperm quality and were not used.

Adult↗

PubMed1994

Predictors of lottery gambling among American college students.

In this study lottery gambling behavior of 288 American college students was examined. Although most students were infrequent gamblers, we found that student lottery gambling was related to having parents and friends who were lottery gamblers. Students who were frequent lottery gamblers were more likely to participate in other forms of gambling and to have begun gambling at younger ages than less frequent gamblers. Locus of control was related to more frequent gambling among parents and only marginally related to more frequent lottery play among students. Discriminant analysis using parental gambling, peer gambling, games played, sex, and locus of control could predict frequency of lottery playing for 72% of gamblers and nongamblers.

Adolescent↗

PubMed1994

Altered gelatinolytic activities in an apparent unilateral keratoconus patient. A case report.

Recent studies have suggested that one possible cause of keratoconus involves increased degradation of the corneal extracellular matrix. Most studies have examined corneas from patients with bilateral, sporadic keratoconus. In this report, both corneas from a 70-year-old man with a history of familial keratoconus and clinical signs of unilateral keratoconus were examined for gelatinase activity. Our results demonstrated a qualitative (three additional activity bands of 88, 92, and 100 kDa molecular weights) and quantitative differences (an apparent increase in overall amounts) in the gelatinolytic enzyme profiles of the affected keratoconus cornea as compared with his unaffected (nonkeratoconus) cornea, normal corneas, bilateral sporadic keratoconus corneas, and cultured keratocytes. In addition, gelatinolytic enzymes were more readily extracted from the affected cornea compared with controls. These data lend support to the hypothesis of a heterogeneous etiology for keratoconus and to previous suggestions that this disorder may be related to an alteration in extracellular matrix degradation.

Adult↗

PubMed1994

Increased gelatinolytic activity in keratoconus keratocyte cultures. A correlation to an altered matrix metalloproteinase-2/tissue inhibitor of metalloproteinase ratio.

Keratoconus is a noninflammatory corneal disorder characterized by gradual stromal thinning and astigmatism. Altered degradation of corneal extracellular matrix is a suggested etiology for this disorder. In the present study we established keratocyte cultures from normal and keratoconus corneas and investigated the roles that matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMP, TIMP-2) may play. After chemical modification (reduction and alkylation) to remove the inhibitor and activation of enzyme with p-aminophenylmercuric acetate (APMA), keratoconus-conditioned media displayed a significant increase (p < 0.05) in the total potential gelatinolytic activity when compared with normal culture media treated in a similar manner. Basal levels of gelatinolytic activity in keratoconus culture media (no reduction, alkylation, or APMA treatment), determined by two different assay methods, tended to be about twice that of normal cell cultures. By zymography, both keratoconus and normal cultures showed identical enzyme patterns, which represented MMP-2 (72 kDa) in its proform and, depending on the treatment of the media, varying amounts of activated MMP-2 (65 kDa). This suggests that the increased gelatinolytic activity in keratoconus was not correlated with an increased appearance of either the 65-kDa-activated form of MMP-2 or a new MMP species. In addition, no differences in the amount of MMP-2 were detected that could account for the increased activities in keratoconus cultures. However, a relative decline in the detectable TIMP levels in keratoconus cultures resulted in an apparent three-fold increase in the ratio of MMP-2/TIMP. Northern blots showed no significant changes in mRNA levels for MMP-1, MMP-2, MMP-3, TIMP, or TIMP-2. These data suggest that a possible alteration in the interaction between MMP-2 and TIMP may play a role in the increased gelatinolytic activity seen in keratoconus tissues.

Adolescent↗

PubMed1994

Clonal lines of Salmonella enterica serotype Enteritidis documented by IS200-, ribo-, pulsed-field gel electrophoresis and RFLP typing.

Sixty-two selected strains of Salmonella serotype Enteritidis of 33 phage types (PTs), and one strain classified as RDNC, were characterised by four different chromosomally based typing methods to elucidate genetic relationships among strains of different phage types. Based on IS200-hybridisation patterns, two major groups, containing strains of the most commonly encountered phage types, and six minor groups (seven with the RDNC strain included) were observed. IS200 pattern was a stable epidemiological marker in strains of all phage types except PT 6a and 14b. Ribotyping separated strains of the phage types into one major and five minor groups; the pattern of the RDNC strain was not seen with other strains. More than one ribotype was observed among strains of Enteritidis PTs 6, 7, 14b and 21. By pulsed-field gel electrophoresis, strains of 21 of the 33 phage types formed one large cluster when bands > 125 kb were used as the criterion for separation. Among strains belonging to PTs 1, 6, 7 and 14b, more than one pattern was observed by this method. By probing with five random cloned fragments of the Enteritidis chromosome, strains from 27 of 31 phage types examined showed the same hybridisation pattern. With the combined use of four genotypic methods, two groups of strains, representing eight and seven of 33 Enteritidis phage types, were formed; these two groups may be considered as the main evolutionary lines of Enteritidis. Strains of the remaining phage types, and the RDNC strain, belonged to separate groups.

Animals↗

PubMed1994

Stability of plasmids in five strains of Salmonella maintained in stab culture at different temperatures.

Four strains of Salmonella berta and one of Salm. enteritidis were stored as stab cultures in sugar-free agar at 5 degrees, 22 degrees and 30 degrees C and in 15% glycerol at -80 degrees C. The stability of the plasmid profiles in each of the strains was monitored over a period of 2.5 years. Plasmid profiles were stable in all strains stored at -80 degrees C, and only six of 450 colonies examined from strains kept in sugar-free agar at 5 degrees C had lost plasmid molecules. Seventy of 440 colonies from stab cultures that were kept at 22 degrees C, and 71 of 440 colonies at 30 degrees C showed changed plasmid profiles. The total number of plasmids lost increased with time, and occasionally, more than one plasmid molecule was lost in the same strain. The virulence associated plasmid of Salm. enteritidis was remarkably stable as it was maintained in all colonies examined at all temperatures investigated. Likewise, no change in SmaI restriction profile was observed in this plasmid molecule at any temperature.

Plasmids↗

PubMed1994

Theileria annulata sporozoite targets.

Bovine peripheral blood mononuclear cells (PBM) infected in vitro with Theileria annulata sporozoites have previously been characterized as MHC class II+ mature macrophages. The ability of T. annulata sporozoites to infect different subpopulations of MHC class II+ bovine monocytes was investigated. Cells were labelled with monocyte specific monoclonal antibodies (MoAb) and isolated using magnetic cell sorting (MACS). Sporozoites infected both immature and mature monocytes, but more readily infected the mature population. A potential ligand for sporozoite entry is the elastin receptor which is expressed mainly on the immature population of monocytes and not on B cells or T cells. T. annulata sporozoites infected elastin receptor positive and negative cell populations equally well. Infected immature cells lost the expression of elastin receptors and the immature marker, subsequently expressing the mature marker. All monocytes lost the expression of CD14 (the LPS receptor) upon infection with sporozoites. The infection of specific populations and subsequent alterations in phenotype may alter the function of these cells and play an important role in disease pathogenesis.

Animals↗

PubMed1994

The characterization of Danish isolates of Salmonella enterica serovar Enteritidis by phage typing and plasmid profiling: 1980-1990.

Plasmid profiling, phage typing and antimicrobial resistance typing have been carried out on 736 isolates of Salmonella enterica serovar Enteritidis collected in Denmark during the period 1980 to 1990. Strains originated from cases of human salmonellosis, broiler poultry flocks, layer poultry flocks, quarantined imported poultry, environmental samples from hatchery units, and from bovines. Phage type (PT) 1 was found to be the most common type among isolates of poultry origin (57.6%) followed by PT4 (28.8%). Isolates belonging to PT8 were found exclusively in imported birds. Phage typing of a representative sample of human isolates revealed the predominance, as in most other Western European countries, of PT4 (61.8%). PT1, however, was found in 17.0% of human strains, a much higher incidence than expected. Antibiotic resistance was observed in 4 out of 107 human isolates (3.7%) and 2 out of 205 non-human isolates (1%) tested.

Animals↗

PubMed1994

Similarities in regulation of the HSV-1 LAT promoter in corneal and neuronal cells.

PURPOSE: To address the possibility of neuronal-like herpes simplex virus type 1 (HSV-1) latency in the cornea by determining if regulation of the HSV-1 LAT promoter in stromal keratocytes is similar to LAT promoter regulation in neurons. METHODS: Transient chloramphenicol acetyltransferase (CAT) assays were used to measure the relative promoter activity of various HSV-1 LAT promoter fragments in primary human corneal cells versus neuronal and nonneuronal cells. RESULTS: The authors found that the LAT promoter, whose location they previously mapped in neurons using transient CAT assays, functioned in stromal keratocytes using the same assay system and that two regions between -283 and -1932 nucleotides relative (upstream) to the start of LAT transcription slightly increased the LAT promoter activity in stromal keratocytes. They previously showed a similar increase in neuronal cells, and a large decrease in nonneuronal cells. In addition, they found that a neuronal specific enhancer region they previously defined between -162 and -283 nucleotides upstream of the start of LAT transcription also enhanced promoter activity in stromal keratocytes. Using gel-shift assays, they detected a nuclear factor specific to neurons and stromal keratocytes that binds to the LAT promoter and that may be a LAT regulatory factor. CONCLUSIONS: Recently, it has been suggested that the cornea might serve as an alternative site of latent herpes simplex virus type 1 (HSV-1) infection. However, this remains controversial. The authors' findings suggest that corneal and neuronal cells regulate the LAT promoter similarly and that this regulation differs from that seen in nonneuronal cells. Thus, the possibility of neuronal-like latency in the cornea remains plausible.

Animals↗

PubMed1993

Whole cell protein profiling of actinobacillus-like strains classified as taxon 2 and taxon 3 according to Bisgaard.

Thirty-nine strains representing all biovars established within the taxa 2 and 3 complex of Bisgaard and two strains belonging to the avian [P.] haemolytical[A.] salpingitidis complex were characterized by one-dimensional SDS-PAGE of cellular proteins. The protein patterns, which contained 40 to 45 discrete protein bands, were highly reproducible. Numerical analysis of the background protein patterns obtained resulted in six major and twelve minor groups (phena). Comparison of the phena defined by protein profiling with species/groups previously established by DNA: DNA hybridization, chemotyping and "biotyping" showed that the best correlation existed between DNA:DNA hybridization and "biotyping". A correlation between results obtained from DNA:DNA hybridization and protein profiling was not obtained. With a few exceptions, a connection was demonstrated between protein profiles and hosts from which the strains belonging to the respective phena originated.

Actinobacillus↗

PubMed1993

Molecular characterization of plasmids in Salmonella enteritidis phage types.

Plasmids in selected type strains of 26 of the Salmonella enteritidis phage types have been characterized by restriction enzyme fingerprinting and by DNA-DNA hybridization with oligonucleotide probes for Salmonella plasmid virulence (Spv) genes. With one exception, the fingerprints of the 38 MDa plasmids studied were homogeneous but there was heterogeneity in the fingerprints of 59 MDa plasmids found in 4 of the type strains. However all 38 MDa and 59 MDa plasmids were related as was a 45 MDa plasmid identified in the type strain of phage type 19. A 3.5 kb fragment homologous to SpvC was conserved in Hind III digests of all 38 MDa and 59 MDa plasmids, and in the related 45 MDa plasmid. In contrast a 65 MDa plasmid found in the type strain of phage type 10 was not related to these three plasmid molecular weight groups and did not carry the SpvC gene.

Base Sequence↗

PubMed1993

Bacterial typing methods suitable for epidemiological analysis. Applications in investigations of salmonellosis among livestock.

The ability to subtype bacteria by typing methods provides the bacteriologist with a powerful means to identify relationships between bacteria. This knowledge is used to identify routes of disease transmission among livestock and from livestock to humans. In the present paper, the principles of bacterial typing and the most commonly applied typing methods for use in veterinary public health are discussed in the context of their application in the investigation of salmonella epidemiology. Typing methods are now routinely used in most investigations on this subject and have provided insight into routes of transmission, reservoirs of infection and mechanisms of persistent infection. Under the EC order on zoonotic diseases, extended surveillance on the presence of zoonotic bacteria in livestock must be expected. To receive the maximum benefits of this surveillance, selected typing methods must be applied to all isolates of e.g. salmonella. At present, serotyping, and phage typing where applicable, are the most obvious choices for continuous surveillance of this organism. Random amplification of polymorphic DNA (PCR based typing) may have the potential for allocating strains into relevant groups quicker and without the requirement for additional manpower, and this method may be preferred in future.

Animals↗

PubMed1993

Susceptibility of transgenic tobacco plants expressing tobacco rattle virus coat protein to nematode-transmitted and mechanically inoculated tobacco rattle virus.

Transgenic Samsun NN tobacco plants expressing the coat protein of tobacco rattle virus were exposed to mechanical leaf inoculation with tobacco rattle virus and to viruliferous trichodorid vector nematodes. Whereas plants were resistant to mechanical inoculation the vector nematodes successfully transmitted tobacco rattle virus to the roots as well as to the leaves of these plants. It is suggested that transgenic resistance is overcome either because vector nematodes inject relatively large numbers of virus particles into a cell or because they inject destabilized particles. The results indicate that coat protein-mediated resistance is unlikely to be of value for controlling tobacco rattle virus in field crops.

Animals↗

PubMed1993

RNA-2 of tobacco rattle virus encodes the determinants of transmissibility by trichodorid vector nematodes.

Pseudorecombinant isolates produced from an efficiently transmitted and a non-transmitted isolate of tobacco rattle tobravirus were tested for their transmissibility by trichodorid vector nematodes. Isolates with RNA-2 originating from the non-transmissible isolate were not transmitted by the vector, whereas isolates with RNA-2 originating from the efficiently transmitted isolate were transmitted efficiently. It is therefore concluded that the factor determining vector transmissibility is located on the RNA-2 genome segment of TRV. Because the serological properties of the isolates also correlate with transmissibility, it is likely that the virus particle protein is involved in the transmission process.

Animals↗