Biomedical subjects
J M Brown
Publications and source records attributed to J M Brown.
Pediatric cardiology.
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Symposium on valvular heart disease.
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Phosphorelay signalling: new tricks for an ancient pathway.
'Two-component' phosphorelay systems, once thought to be used exclusively by prokaryotes, have in the past few years been shown to exist in eukaryotes. A two-component system in Dictyostelium has now been shown to modulate protein kinase A, a key regulator of multicellular development in this organism.
lagC-null and gbf-null cells define key steps in the morphogenesis of Dictyostelium mounds.
The transition to multicellularity is a key feature of the Dictyostelium life cycle, and two genes, gbf and lagC, are known to play pivotal roles in regulating this developmental switch. lagC-null and gbf-null cells fail to induce cell-type-specific genes ordinarily expressed during multicellular development. The null mutants also share a similar morphological phenotype: mutant cells repeatedly aggregate to form a loose mound, disperse, and reform a mound, rather than proceeding to form a tip. To characterize defects in morphogenesis in these mutants, we examined cell motion in the mutant mounds. In analogy with the failed transition in gene expression, we found that lagC-null and gbf-null mounds failed to make a morphogenetic transition from random to rotational motion normally observed in the parent strain. One reason for this was the inability of the mutant mounds to establish a single, dominant signaling-wave center. This defect of lagC-null or gbf-null cells could be overcome by the addition of adenosine, which alters cAMP signaling, but then even in the presence of apparently normal signaling waves, cell motility was still aberrant. This motility defect, as well as the signaling-wave defect, could be overcome in lagC-null cells by overexpression of GBF, suggesting that lagC is dispensable if GBF protein levels are high enough. This set of morphogenetic defects that we have observed helps define key steps in mound morphogenesis. These include the establishment of a dominant signaling-wave center and the capacity of cells to move directionally within the cell mass in response to guidance cues.
Tirapazamine is metabolized to its DNA-damaging radical by intranuclear enzymes.
Tirapazamine (TPZ), a new anticancer drug that is currently in Phase II and III clinical trials, has a unique mechanism of action. Its cytotoxicity is selective for hypoxic cells in solid tumors and results from DNA damage produced by a free radical, which is generated by enzymatic reduction of the parent molecule. However, there is no agreement as to which enzyme(s) are involved. Here, we have measured both DNA damage and TPZ metabolism in A549 human lung cancer cells and in isolated nuclei derived from the cells. We show that, although the nuclei metabolize TPZ at a rate that is only 20% of that of whole cells, they have levels of DNA damage that are similar to those of the cells. We also show that TPZ radicals that are formed outside nuclei do not contribute to intranuclear DNA damage. Thus, essentially all of the DNA damage from TPZ results from radicals generated within the nucleus, and the 80% of the drug metabolism that occurs in the cytoplasm is probably irrelevant for the activity of this drug in killing hypoxic cells.
The unique physiology of solid tumors: opportunities (and problems) for cancer therapy.
The physiology of solid tumors differs from that of normal tissues in a number of important aspects, the majority of which stem from differences between the two vasculatures. Compared with the regular, ordered vasculature of normal tissues, blood vessels in tumors are often highly abnormal, distended capillaries with leaky walls and sluggish flow. Tumor growth also requires continuous new vessel growth, or angiogenesis. These physiological differences can be problems for cancer treatment; for example, hypoxia in solid tumors leads to resistance to radiotherapy and to some anticancer drugs. However, these differences can also be exploited for selective cancer treatment. Here we review four such areas that are under active investigation: (a) hypoxia-selective cytotoxins take advantage of the unique low oxygen tension in the majority of human solid tumors. Tirapazamine, a drug in the final stages of clinical trials, is one of the more promising of these agents; (b) leaky tumor blood vessels can be exploited using liposomes that have been sterically stabilized to have a long intravascular half-life, allowing them to selectively accumulate in solid tumors; (c) the tumor microenvironment is a stimulus to angiogenenesis, and inhibition of angiogenesis can be a powerful anticancer therapy not susceptible to acquired drug resistance; and (d) we discuss attempts to use gene therapy activated either by the low oxygen environment or by necrotic regions of tumors.
Cancer therapy and tumor physiology.
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Muscles within muscles: the neuromotor control of intra-muscular segments.
The aim of this investigation was to anatomically identify, and then determine the function of, individual segments within the human deltoid muscle. The anatomical structure of the deltoid was determined through dissection and/or observation of the shoulder girdles of 11 male cadavers (aged 65-84 years). These results indicate that the deltoid consists of seven anatomical segments (D1-D7) based upon the distinctive arrangement of each segment's origin and insertion. Radiographic analysis of a cadaveric shoulder joint suggested that only the postero-medial segment D7 has a line of action directed below the shoulder joint's axis of rotation. The functional role of each individual segment was then determined utilising an electromyographic (EMG) technique. Seven miniature (1 mm active plate; 7 mm interelectrode distance) bipolar surface electrodes were positioned over the proximal portion of each segment's muscle belly in 18 male and female subjects (18-30 years). EMG waveforms were then recorded during the production of rapid isometric shoulder abduction and adduction force impulses with the shoulder joint in 40 degrees of abduction in the plane of the scapula. Each subject randomly performed 15 abduction and 15 adduction isometric force impulses following a short familiarisation period. All subjects received visual feed back on the duration and amplitude of each isometric force impulse produced via a visual force-time display which compared subject performance to a criterion force-time curve. Movement time was 400 ms (time-to-peak isometric force) at an intensity level of 50% maximal voluntary contraction. Temporal and intensity analyses of the EMG waveforms, as well as temporal analysis of the isometric force impulses, revealed the neuromotor control strategies utilised by the CNS to control the activity of each muscle segment. The results showed that segmental neuromotor control strategies differ across the breadth of the muscle and that individual segments of the deltoid can be identified as having either "prime mover", "synergist", "stabiliser" or "antagonist" functions; functional classifications normally associated with whole muscle function. Therefore, it was concluded that the CNS can "fine tune" the activity of at least six discrete segments within the human deltoid muscle to efficiently meet the demands of the imposed motor task.
Genetic targets for the detection and identification of Venezuelan equine encephalitis viruses.
Rt-PCR probes targeted to different gene sequences of VEE (Venezuelan equine encephalitis) virus strain TC-83 were assessed for their sensitivity, specificity and non-specific cross-reactivity. A generic VEE virus amplimer (VNSP4F2/VNSP4R2), targeted against nsP4 was identified, which was sensitive (detected at least 10 pfu) and robust (worked over a wide range of salt concentrations and annealing temperatures). An E2 amplimer designed against TC-83, (VE2F/VE2R), identified VEE strains TRD (1AB), P676 (1C), 3880 (1D) Everglades (2) vRNA whilst a second E2 primer pair designed against strain 68U201, (68UF/68UR), identified all the remaining VEE viruses in the sero-complex. This would suggest that the VEE virus E2 gene can be sub-divided at the genetic level into two separate groups making it a useful target for differentiation of serosubtypes 1 and 2 from the other VEE virus subtypes. Using a panel of amplimers targeted to different VEE genes and strains it was possible to distinguish between most of the serotypes, but most importantly, we were able to detect the epizootic strains TRD and P676 as well as other VEE viruses implicated in human disease (sero-subtypes 1D and 1E).
Resection of a leiomyosarcoma of the azygos vein.
We report the resection of a leiomyosarcoma of the azygos vein in an elderly woman. The rare nature of the tumor and the unusual site of origin are discussed.
Acute and chronic effects of bilateral lung transplantation without cardiopulmonary bypass on the first transplanted lung.
BACKGROUND: Bilateral lung transplantation (BLT) without cardiopulmonary bypass (CPB) may exacerbate reperfusion injury to the initially engrafted lung because of increases in pulmonary flow during implantation of the second graft. METHODS: In a retrospective review of 23 BLT patients, we hypothesized that BLT without CPB injures the first transplanted lung measured by acute and late graft dysfunction compared to the second transplanted lung. Of the 23 BLT, 19 underwent transplantation without CPB while 4 patients were placed on CPB secondary to hemodynamic instability. RESULTS: Acute graft function was assessed by radiographic scoring of lung quadrants (blinded radiologist; 0 = no infiltrate; 1 = infiltrate; maximum = 2 per lung) and by arterial/alveolar oxygen tension ratios (PaO2/ FiO2) ratios. Late graft function was evaluated by quantitative perfusion scan. Lung perfusion was graded as abnormal if less than 50% on the right or less than 45% on the left (Fisher's exact). Radiographic scores were not different between first and second implanted lungs at 1 and 24 hours, PaO2/FiO2 ratios at 1 and 24 hours were 273+/-26 and 312+/-23, respectively, and perfusion scans at 3 and 12 months revealed normal differential blood flow. CONCLUSIONS: These findings suggest no acute or chronic differences occur between the first or second transplanted lung completed without CPB.
Geographic variations in the composition of ivory of the African elephant (Loxodonta africana).
Tracing the source of origin of illegal ivory will contribute to the identification of poorly managed game parks and facilitate steps taken to prevent the African elephant from becoming extinct. This study was aimed at establishing a database on the composition of ivory obtained from elephant sanctuary areas in Southern Africa. Fragments of elephant ivory from seven geographically distinct areas in South Africa, Namibia and Botswana were analysed for inorganic and organic content. A total of 20 elements was detected in the inorganic fraction of ivory, some in concentrations as low as 0.25 microg/g. The concentrations of calcium, phosphate, magnesium, fluoride, cobalt and zinc showed statistically significant differences (p < 0.007) between ivory obtained from different regions. Analyses of the organic fraction identified 17 amino acids. Ivory from arid regions showed significantly lower proline plus hydroxyproline content and under-hydroxylation of lysine residues. This study indicates that chemical analyses of ivory could be beneficial in tracing the source of illegal ivory.
Modified ice study: a phase II study of an intensive, modified ICE regimen (ifosfamide, carboplatin and etoposide) in patients with better prognosis, small cell lung cancer.
A total of 30 with good prognosis small cell lung cancer were treated with a modified 'ICE' (ifosfamide, carboplatin and etoposide) chemotherapy regimen in an attempt to achieve a high response rate with less toxicity than is seen with the full 'ICE' regimen. This was given every 4 weeks for a maximum of six cycles. In total 25 patients (83%, 95% CI (70-97%)) experienced a partial or complete response at some stage of their treatment. Of these patients, 12 (40%, 95% CI (22-58%)) showed a complete response. A total of 19 patients (63%) had to have their dose reduced and/or delayed at some point due to toxicity. Nadir blood counts showed that 19 patients (63%, 95% CI (46-81%)) had WHO grade 3 or 4 thrombocytopenia, and 24 (86%, 95% CI (73-99%)) had grade 3 or 4 neutropenia. A total of 17 patients (53%) completed six cycles of chemotherapy. In total 3 patients died during treatment all due to treatment-related complications. Median survival was 12.6 months (95% CI (11.6, 14.7 months)). Nausea, vomiting, dysphagia, activity, mood and overall condition, as recorded using daily diary cards, were worse at the beginning of each chemotherapy cycle. Both response rates and survival were clinically acceptable. However, neutropenia and thrombocytopenia, although reduced from rates reported with the full ICE regimen, were still high. A prospective randomised controlled trial is now needed to assess this regimen in more detail.
Contrast-enhanced sonography of tumor neovascularity in a rabbit model.
This project was designed to detect the development of tumor neovascularity and determine if intravenous microbubble contrast improves visualization of otherwise undetectable tumors in an animal model. VX-2 carcinoma was implanted into one thigh of 10 New Zealand white rabbits. Tumors were assessed without and with contrast at 1- to 4-day intervals from day 3-19 postimplantation, using gray scale, color flow, pulse Doppler and power Doppler imaging. Tumor vascularity was compared with the contralateral thigh muscle, so each animal was its own control. Contrast injection improved visualization of tumor neovascularity. Early tumors had homogeneous vasculature but, with time, the centers became less vascular, while the periphery increased. Following contrast injection, color gain was decreased by 40% without compromising color intensity. Neovascularity was detected by contrast injection before the tumor could be palpated or visualized by gray scale. Based on these data, we conclude that enhancement of neovascularity by intravenous contrast permits earlier detection and improved visualization of soft tissue tumors in rabbits.
Lower extremity volumetric arterial blood flow in normal subjects.
The objective of this clinical study was to establish normal values for volumetric blood flow in the leg at rest using Doppler ultrasound, and to determine what biophysical factors influence resting volumetric flow. Arterial blood flow was measured at four sites in the legs of 40 healthy subjects using an ATL Ultramark 9 HDI system. All subjects were nonhypertensive nonsmokers with ankle brachial index values greater than 1 and no history of vascular disease. The subjects, 20 of each gender, in age ranging from 20 to 64 y were examined. Blood flow was calculated from the time-averaged, intensity-weighted mean velocity Doppler waveforms and vessel cross-sectional area at the same site. Thigh and calf circumference measurements were used to estimate muscle masses. The mean flow and standard error measured in four arteries in the leg were: 284+/-21 mL/min in the common femoral (CFA); 152+/-10 mL/min in the superficial femoral (SFA); 72+/-5 mL/min in the popliteal; and 3+/-1 mL/min in the dorsalis pedis. Although women tended to have higher time-averaged mean velocities in the CFA and SFA than men (t-test, p < 0.008), their arterial cross-sectional areas tended to be smaller (t-test, p < 0.004) and no statistically significant difference was found between men and women in volumetric flow at any site. No correlation was found between age, weight, height, muscle mass and volumetric flow at all four sites. These estimates of lower extremity volumetric flow in healthy subjects provide a baseline for future studies of flow rates in patients with vascular disease.
Cryomyolysis, a new procedure for the conservative treatment of uterine fibroids.
Conservative surgical options for uterine myomata traditionally were abdominal myomectomy, laparoscopic myomectomy, and, more recently, myolysis. Each of these procedures has distinct advantages, but also apparent disadvantages. We attempted to introduce an additional option for conservative surgical treatment of fibroids by freezing the structures, a procedure termed cryomyolysis. In this pilot study, 14 women were pretreated with a gonadotropin-releasing hormone (GnRH) agonist for a minimum of 2 months preoperatively to minimize uterine and myoma size. Cryomyolysis was performed and the GnRH agonist was discontinued. Magnetic resonance imaging scans were performed in 10 of the 14 women after GnRH agonist treatment but before surgery, and 4 months postoperatively. Total uterine volume ranged from 41.3 to 1134.8 ml preoperatively, and 49.5 to 1320 ml postoperatively (mean increase 22% after discontinuation of GnRH agonist). Normal uterine volume ranged from 35.6 to 548.7 ml preoperatively and 45.1 to 729.6 ml postoperatively (mean increase 40%); however, myoma volume showed a mean decrease of 6% (range -87-28%). Analysis of only frozen myomata revealed a mean volume decrease of 10%. Cryomyolysis maintains at or slightly reduces these lesions to post-GnRH agonist size, and all other uterine tissue returns to pretreatment size. We believe cryomyolysis may be an effective conservative surgical approach to uterine fibroids.
Exploiting tumour hypoxia and overcoming mutant p53 with tirapazamine.
Human solid tumours are composed of a significant proportion of hypoxic cells, i.e. cells with oxygen levels lower than those of normal tissues. Tumour hypoxic cells have been shown to have a negative impact on the response of solid tumours to radiation therapy and chemotherapy. However, these low cellular oxygen levels can be exploited by a drug that is specifically activated to a cytotoxic metabolite at these low levels. Tirapazamine is a novel bioreductive agent with selective cytotoxicity to hypoxic tumour cells, irrespective of their p53 status or apoptotic response, and acts synergistically with cisplatin. This potentiation is dependent on an interaction that can only take place in a hypoxic environment, resulting in a significant sensitization of the cells to cisplatin cell killing, with no increase in the systemic toxicity of cisplatin. Thus, the low cellular oxygen levels common in solid tumours can be turned from disadvantage to advantage using the hypoxia-selective cytotoxic drug tirapazamine.