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Biomedical subjects

D Phillips

Publications and source records attributed to D Phillips.

At least 73 records · Page 4Linked to original sources

Laser line-scanning confocal fluorescence imaging of the photodynamic action of aluminum and zinc phthalocyanines in V79-4 Chinese hamster fibroblasts.

Confocal fluorescence microscopy, using a newly constructed laser line-scanning confocal microscope, was applied to an investigation of the early stages of photoinduced destruction of V79-4 Chinese hamster fibroblasts using aluminum and zinc phthalocyanines as photosensitizers. Results obtained in this work show that aluminum and zinc phthalocyanines, once internalized, localize in perinuclear sites that are disrupted upon light exposure resulting in fluorescence redistribution. The combination of laser-line scanning with charge-coupled device detection used in the confocal microscope developed in this work can enable rapid high-resolution sequential imaging, which is ideal for studying photoinduced intracellular fluorescence dynamics.

Animals↗

Effect of gonadotrophin and sex steroid on the scotoresponses of day old chicks of Japanese quail, Coturnix coturnix japonica.

Luteinizing hormone (LH) and testosterone propionate (TP) were tested for their effects on the development of scotorefractoriness in Japanese quail. Day old chicks of both the sexes were divided into four groups and treated with normal saline, LH (1 microg/100 g body weight) and two doses of TP (100 microg/100 g body weight- TP1 and 1 mg/100 gm body weight- TP2) over a periods of 14 weeks. In the male chicks, compared to control, LH treatment advanced and low dose of TP suppressed the development of scotorefractoriness, while high dose of TP inhibited it completely and maintained the birds in scotosensitive state. On the other hand, in females, LH treatment increased the rate of sexual development resulting in the onset of egg laying earlier than that of control, but both the doses of steroid hormone suppressed ovarian development. It is suggested that, LH treatment not only induced a higher degree of reproductive development in short day quail but may also advance sexual maturity as under long daylength. Further, both the doses of male hormone had negative feedback effect on neuroendocrine axis and eliminated the attainment of scotorefractoriness i.e., reproductive development under short days.

Animals↗

Ligamentomuscular protective reflex in the elbow.

A reflex are from the medial elbow ligaments to the forearm pronator muscles was shown to exist in the feline model. A single articular branch emerging from the median nerve and converging on the medial collateral ligament was identified and stimulated with supramaximal pulses of 100 microseconds duration at a rate of 10 pulses/s. Stimulation of the articular nerve elicited myoelectric activity in the flexor digitorum superficialis, flexor digitorum profundus, flexor carpi radialis, flexor carpi ulnaris, and pronator teres. Transection of the articular nerve between the electrodes and the median nerve resulted in the disappearance of any myoelectric activity in the muscles, thus confirming the afferent nature of the articular nerve. The mean time delay from the application of the stimulus to the corresponding myoelectric discharge ranged from 3.2 to 5.8 ms for the 5 muscles. The existence of a fast-acting reflex arc from the medial elbow ligaments to the forearm muscles both confirms the concept of ligamentomuscular protective synergy (shown to exist in the knee, shoulder, and ankle joints) and extends it to the elbow. This reflex arc has significant implications for both the planning of elbow surgery while preserving the neural supply of the ligaments and for the planning of postsurgical or conservative therapeutic rehabilitation modalities.

Afferent Pathways↗

Excited triplet state photophysics of the sulphonated aluminium phthalocyanines bound to human serum albumin.

The binding of the sulphonated aluminum phthalocyanines to human serum albumin (HSA) in aqueous phosphate buffer solution at 25 degrees C has been studied by measuring the properties of the triplet excited states of these dyes. The triplet lifetimes were measured by triplet-triplet absorption flash photolysis. The triplet lifetime of the disulphonated AlS2Pc (2.5 microM) varies from 500 +/- 30 microseconds in the absence of protein to 1.100 microseconds and longer with HSA concentrations above 100 microM. Under identical conditions, the maximum triplet lifetimes of the mono-, tri- and tetrasulphonated compounds bound to HSA are shorter than those for the disulphonated species. The increase in the triplet state lifetimes is attributed to the ability of the bulk aqueous phase to interact with the sensitizer at the site of binding; the site of binding being dependent on the degree of sulphonation. For AlS2Pc and AlS3Pc at all HSA concentrations, and regardless of the degree of sulphonation, all the triplet state decay profiles follow simple pseudo-first-order kinetics. The exponential decay of the triplet phthalocyanine at all HSA concentrations is ascribed to the rapid association and dissociation of the phthalocyanine-HSA complex on the time-scales of the triplet state lifetimes. A simplified one-step binding model is utilized to describe the results. The association of AlS1Pc with HSA results in substantial quenching of the triplet state quantum yield, and a more complex model is required to analyze the results. The tetrasulphonated compound (AlS4Pc) binds to the protein at a site where it experiences some protection from the aqueous phase.

Aluminum↗

A hydroxypyridinone (CP94) enhances protoporphyrin IX formation in 5-aminolaevulinic acid treated cells.

Different cell lines were given photodynamic treatment with 5-aminolaevulinic acid (ALA) and light. In addition, the iron chelator 1,2-diethyl-3-hydroxypyridin-4-one (CP94) was used. The porphyrin species produced was spectrofluorimetrically identified as protoporphyrin IX. All the cell lines responded to treatment, including a multidrug resistance gene expressing bladder cancer line and, to a lesser degree, cells derived from untransformed human skin fibroblasts. CP94 enhanced both porphyrin fluorescence, total porphyrin content and photosensitivity of the cells. CCD fluorescence microscopy showed a granular extranuclear porphyrin fluorescence distribution for all the cell lines involved, but the untransformed cells showed a distribution pattern different from the ones seen in the other cells.

Aminolevulinic Acid↗

Idoxifene is equipotent to tamoxifen in inhibiting mammary carcinogenesis but forms lower levels of hepatic DNA adducts.

Tamoxifen is an effective agent preventing mammary carcinogenesis in rats but causing liver tumours. Idoxifene is a more potent antioestrogen and is effective in patients with advanced breast cancer. We therefore compared the effects of idoxifene with tamoxifen on mammary carcinogenesis and hepatic DNA adduct formation. To do this, we undertook a study designed to compare tamoxifen with idoxifene as a chemopreventive agent in rats inoculated with N-methylnitrosourea (MNU) and also measured hepatic adduct formation. We examined the time to mammary tumour development in 272 female Ludwig/Wistar/Olac rats treated with MNU followed by tamoxifen (5 mg kg(-1)), equimolar idoxifene or vehicle three times a week for up to 24 weeks. To determine duration of effect, a second study was carried out whereby all of the 129 animals surviving at the end of treatment were entered into a surveillance programme for 27 weeks after the end of the administration period. Hepatic DNA adduct formation was examined by 32P-postlabelling in a group of rats after 24 weeks' treatment. In the first study, both idoxifene and tamoxifen were effective in preventing tumour growth as only 2 out of 21 (10%) MNU and vehicle-treated animals were alive and tumour free after 24 weeks compared with 13 out of 22 (59%) animals receiving MNU followed by idoxifene or tamoxifen (P < 0.001). The second study showed that, in both idoxifene- and tamoxifen-treated animals, a progressive regrowth of tumours occurred after cessation of therapy, as by the end of the observation period only four idoxifene-treated animals and one tamoxifen-treated animal were free from disease. In the subset of animals tested, tamoxifen-treated animals had approximately 100 times higher levels of DNA hepatic adducts than idoxifene-treated animals. Adducts were not seen in the control group. These results indicate that idoxifene is as effective a chemopreventive agent as tamoxifen in the rat while causing only very low levels of DNA adducts in liver tissue and suggest that idoxifene may be a well-tolerated chemopreventive agent in women who are at increased risk of breast cancer.

Animals↗

Comparative photophysical study of disulfonated aluminum phthalocyanine in unilamellar vesicles and leukemic K562 cells.

The photophysical properties of cis-disulfonated aluminum phthalocyanine (AlPcS2) in unilamellar vesicles (liposomes) of DL-alpha-dipalmitoyl-phosphatidylcholine have been measured. Both the fluorescence and triplet quantum yields decreased with increasing sensitizer concentration. The time-resolved fluorescence decays, analyzed by both the sum of exponentials and decay time distribution analyses, are compared with those reported for AlPcS2 in leukemic K562 cells. Information on the photodynamic transport and localization mechanism has been obtained by drawing correlations between the two systems, indicating active transport of the phthalocyanine into tumor cells involving lysosomal accumulation.

1,2-Dipalmitoylphosphatidylcholine↗

Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization.

We have developed a new system of chromosomal mutagenesis in order to study the functions of uncharacterized open reading frames (ORFs) in wild-type Escherichia coli. Because of the operon structure of this organism, traditional methods such as insertional mutagenesis run the risk of introducing polar effects on downstream genes or creating secondary mutations elsewhere in the genome. Our system uses crossover PCR to create in-frame, tagged deletions in chromosomal DNA. These deletions are placed in the E. coli chromosome by using plasmid pKO3, a gene replacement vector that contains a temperature-sensitive origin of replication and markers for positive and negative selection for chromosomal integration and excision. Using kanamycin resistance (Kn(r)) insertional alleles of the essential genes pepM and rpsB cloned into the replacement vector, we calibrated the system for the expected results when essential genes are deleted. Two poorly understood genes, hdeA and yjbJ, encoding highly abundant proteins were selected as targets for this approach. When the system was used to replace chromosomal hdeA with insertional alleles, we observed vastly different results that were dependent on the exact nature of the insertions. When a Kn(r) gene was inserted into hdeA at two different locations and orientations, both essential and nonessential phenotypes were seen. Using PCR-generated deletions, we were able to make in-frame deletion strains of both hdeA and yjbJ. The two genes proved to be nonessential in both rich and glucose-minimal media. In competition experiments using isogenic strains, the strain with the insertional allele of yjbJ showed growth rates different from those of the strain with the deletion allele of yjbJ. These results illustrate that in-frame, unmarked deletions are among the most reliable types of mutations available for wild-type E. coli. Because these strains are isogenic with the exception of their deleted ORFs, they may be used in competition with one another to reveal phenotypes not apparent when cultured singly.

Alleles↗

Early infection and subsequent insulin dependent diabetes.

In a study of 58 children under the age of 16 with insulin dependent diabetes (IDDM) and 172 matched non-diabetic controls, infection during the first year of life was associated with a reduction in diabetes risk (odds ratio 0.81, 95% confidence interval 0.67 to 0.98, per infective episode). Decreased exposure to common infections during infancy may be linked with subsequent IDDM.

Adolescent↗

Isolation of Japanese encephalitis virus from Culex annulirostris in Australia.

Mosquitoes were collected using CO2 and octenol-baited light traps during an outbreak of Japanese encephalitis (JE) on Badu Island in the Torres Strait, Australia in April 1995. A total of 13,300 mosquitoes comprising 12 species were processed for virus isolation. Eight isolates of JE virus were obtained from Culex annulirostris, with a carriage rate of 2.97:1,000; this mosquito also yielded one Sindbis virus isolate. A reverse transcriptase-polymerase chain reaction was used to sequence the JE viruses, which were compared with published sequences. The eight isolates were 90% homologous with known JE strains but only 68% homologous with other flaviviruses. Among the isolates, 99% homology was obtained, suggesting a common point of origin.

Aedes↗

Claudication distance is poorly estimated and inappropriately measured.

BACKGROUND: Claudication distance is the commonest measure of the disability caused by lower-limb occlusive arterial disease. The accuracy of claudication distance as a surrogate for handicap has been assessed. METHODS: Seventy patients who attended a specialist vascular clinic with intermittent claudication were studied prospectively. Patients were asked to estimate their claudication distance and maximum walking distance before undergoing both a patient-controlled corridor walk and a fixed-speed treadmill walk. RESULTS: The claudication distance reported by patients bore little relation to the distance recorded in the medical correspondence. There was no correlation between the estimated distance and the actual distance walked on either a patient-controlled corridor walk or a fixed-speed treadmill walk. Most patients were able to walk substantially further at their own speed on the corridor than on the treadmill at a slower speed. CONCLUSION: Claudication distance is spuriously estimated, inaccurately reported, falsely recorded, inappropriately measured and usually misinterpreted. It is of little value in judging the need for treatment. Objective measures of the handicap caused by the disability of reduced walking distance are required if rational management decisions are to be made.

Adult↗

Characterization of monoclonal antibodies to Pseudomonas aeruginosa type A flagellar antigen.

Flagellar murine MAbs (53B and 29) to strain a-type 170018 (a0,a3,a4, 45 kDa) and a human a-type MAb were studied in ELISA, Immunoblot, colony blot, agglutination and motility assays to evaluate the degree of cross-reactivity within the dominant a0 epitope. No specific effect of possible subtypes (a1, a2, a3, a4) was observed. An association of cross-reactivity and molecular weight was observed for 53B. A broader cross-reactivity as seen with MAbs 29 and A522 including high molecular weight flagellins (49-52 kDa), and particularly in the motility assay, may predict protective potential. Moderate reactivity with strain 5939 (a0 a3) was only seen with A522 MAb. These data indicate the presence of several cross-reactive sites associated with the common a0 antigen.

Agglutination Tests↗

Wound cleaning versus wound disinfection: a challenging dilemma.

Our experience with this resident has had a significant impact on our approach to wound management. We no longer accept normal saline as the only option for wound cleansing. Instead, we approach wound cleansing systematically using the wound cleansing model outlined in Figure 1. If healing is not apparent, we critically review the situation and consider the bacterial status of the wound, the phase of wound healing, and the effects of wound cleansing versus wound disinfection on both the bacteria and the cells responsible for wound repair. In infected wounds or those colonized with a high bacterial count, careful attention is given to eradicating bacterial contamination. We carefully weigh the benefits of wound disinfection (eradication of bacterial contamination) against wound cleansing and the potential harm to the resident (delayed wound healing and unnecessary discomfort). If the benefits of wound disinfection outweigh the potential harm to the resident, we choose wound disinfection and monitor its effectiveness on a daily basis. When bacterial contamination has been eradicated and the wound is clean, we resume wound cleansing with normal saline.

Acetic Acid↗

Distress and radiological investigations of the urinary tract in children.

UNLABELLED: The distress associated with radiological investigations may be related to the information and style of parental explanation given to the child and the parents own coping style. In a questionnaire survey we assessed distress in relation to investigations of the urinary tract using the Groningen Distress Rating Scale and the Utrecht Coping List. We found the micturating cystourethrogram (MCUG) to be significantly more distressing than radionuclide investigations. MCUG distress was adversely correlated with passive and palliative parental coping styles. Results suggested that cognitive coping styles modelled by the parents ameliorated distress on dimercaptosuccinic acid scintigraphy. CONCLUSION: The high level of MCUG associated distress and presence of traumatised children in the MCUG sample, along with the potential for parental contribution to distress reduction, suggests that both children and parents would benefit from more active preparation programmes prior to MCUG testing.

Adaptation, Psychological↗