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Biomedical subjects

J A Green

Publications and source records attributed to J A Green.

At least 19 recordsLinked to original sources

Estimating the rate of oxygen consumption during submersion from the heart rate of diving animals.

How animals manage their oxygen stores during diving and other breath-hold activities has been a topic of debate among physiologists for decades. Specifically, while the behavior of free-ranging diving animals suggests that metabolism during submersion must be primarily aerobic in nature, no studies have been able to determine their rate of oxygen consumption during submersion (Vo(2)d) and hence prove that this is the case. In the present study, we combine two previously used techniques and develop a new model to estimate Vo(2)d accurately and plausibly in a free-ranging animal and apply it to data for macaroni penguins (Eudyptes chrysolophus) as an example. For macaroni penguins at least, Vo(2)d can be predicted by measuring heart rate during the dive cycle and the subsequent surface interval duration. Including maximum depth of the dive improves the accuracy of these predictions. This suggests that energetically demanding locomotion events within the dive combine with the differing buoyancy and locomotion costs associated with traveling to depth to influence its cost in terms of oxygen use. This will in turn effect the duration of the dive and the duration of the subsequent recovery period. In the present study, Vo(2)d ranged from 4 to 28 ml.min(-1).kg(-1), indicating that, at least as far as aerobic metabolism was concerned, macaroni penguins were often hypometabolic, with rates of oxygen consumption usually below that for this species resting in water (25.6 ml.min(-1).kg(-1)) and occasionally lower than that while resting in air (10.3 ml.min(-1).kg(-1)).

Animals↗

Morphologic and histologic comparisons between in vivo and nuclear transfer derived porcine embryos.

Nuclear transfer (NT) is an inefficient but invaluable tool of the biotechnology industry. This study looked at abnormalities associated with peri-implantation NT porcine embryos. Four experimental groups were examined: nonpregnant animals, in vivo pregnant animals, NT recipients, and manipulation control embryos (MC). Embryos (Day 10, 12, or 14) were evaluated for embryonic disc diameter, gross morphology, nucleoli density, and mitotic figure index. Day 12 (P < or = 0.03) and Day 14 (P < or = 0.01) NT embryos had increased numbers of nucleoli, and Day 14 NT embryos had an increased (P < or = 0.03) mitotic index compared to in vivo and MC embryos. In vivo produced Day 14 embryos had increased (P < or = 0.01) disk diameters when compared to other embryos except for MC Day 14, which also showed increases (P < or = 0.01) in disk diameter except when compared to in vivo produced Day 12 and Day 14 embryos. In vivo produced Day 12 had greater (P < or = 0.03) disk diameters when compared to NT and MC embryos except for MC Day 14, and in vivo produced Day 14 embryos, which had a significantly increased (P < or = 0.01) disk diameter. In vivo produced Day 14 embryos were morphologically more advanced (P < or = 0.01) than Day 14 NT and MC counterparts. NT embryos develop at a slower rate than their in vivo produced counterparts. The increase in nucleoli and mitotic index of NT embryos suggest the cell cycle may be affected or the NT embryos are employing other means to compensate for slow development. The techniques used during NT also appear to compromise embryo development.

Animals↗

Chemotherapy for advanced, recurrent or metastatic endometrial cancer: a systematic review of Cochrane collaboration.

BACKGROUND: Cytotoxic chemotherapy has a limited place in the management of advanced or recurrent endometrial cancer. Commonly used agents include cisplatin and doxorubicin, but the side-effect profile may be unacceptable for many patients. The feasibility of administration of combination chemotherapy is limited in many patients on account of significant co-morbidity. While early-stage endometrial adenocarcinoma is a common gynaecological cancer with a favourable prognosis, advanced or recurrent disease presents a difficult management problem. The platinum and anthracycline compounds have been widely used for many years, but their impact on progression-free survival (PFS) and overall survival (OS) is not clear. This systematic review aimed to evaluate both the benefits and adverse effects of cytotoxic chemotherapy in these women. PATIENTS AND METHODS: We carried out systematic searches for randomised controlled trials (RCTs) comparing chemotherapy with another intervention. Data were extracted from trial reports or supplied by investigators. Where possible, hazard ratios (HRs) were calculated for OS and PFS and odds ratios (ORs) were calculated for acute toxicity. The impact of more versus less intensive chemotherapy on OS, PFS and acute toxicity was assessed in a meta-analysis. RESULTS: Eleven eligible RCTs were identified that recruited 2288 patients. A meta-analysis of six of these trials found that PFS [HR = 0.80, 95% confidence interval (CI) 0.71-0.90; P = 0.004], but not OS (HR = 0.90, 95% CI 0.80-1.03; P = 0.12), was significantly improved when more intensive chemotherapy was compared with less intensive chemotherapy. OS was improved when doxorubicin, cisplatin and other drugs were compared with doxorubicin and cisplatin. Toxicity was generally higher with more chemotherapy. There was insufficient evidence to assess the effect of chemotherapy on symptom control or quality of life (QoL). Platinums, anthracyclines and taxanes were the most studied in phase II trials and combinations gave the best responses, but patient selection and pre-treatment was very variable. CONCLUSIONS: More intense combination chemotherapy significantly improves the disease-free survival and the data indicate a modest improvement in OS. The addition of anthracyclines (e.g. doxorubicin) or the taxanes [e.g. paclitaxel (Taxol)] to cisplatin increases the response rate. More intensive regimens are associated with the gain in survival. However, grade 3 and 4 myelosuppression and gastrointestinal toxicity are also increased. Future developments are likely to exploit specific molecular characteristics of endometrial cancers, including their hormone dependence, growth factor target overexpression and PTEN loss. While no one drug or regimen offers a clear benefit for women with advanced endometrial cancer, platinum drugs, anthracyclines and paclitaxel seem the most promising agents. Future trials should address the impact of such agents on QoL and symptom control in addition to survival. Chemotherapy and endocrine therapy need to be compared directly in an RCT.

Antineoplastic Agents↗

Physiological response to feeding in little penguins.

Specific dynamic action (SDA), the increase in metabolic rate above resting levels that accompanies the processes of digestion and assimilation of food, can form a substantial part of the daily energy budget of free-ranging animals. We measured heart rate (fH) and rate of oxygen consumption (VO2) in 12 little penguins while they digested a meal of sardines in order to determine whether they show specific dynamic action. In contrast to some studies of other penguin species, little penguins showed a substantial SDA, the magnitude of which was proportional to the size of the meal. The energy utilized in SDA was equivalent to 13.4% of the available energy content of the fish. Furthermore, animals such as penguins that forage in a cold environment will probably expend further energy in heating their food to body temperature to facilitate efficient digestion. It is estimated that this additional energy expenditure was equivalent to 1.6%-2.3% of the available energy content of the fish, depending on the time of year and therefore the temperature of the water. Changes in fH during digestion were qualitatively similar to those in VO2, implying that there were no substantial circulatory adjustments during digestion and that the relationship between fH and VO2 in penguins is unaffected by digestive state.

Animals↗

Alterations in the p53 pathway and prognosis in advanced ovarian cancer: a multi-factorial analysis of the EORTC Gynaecological Cancer group (study 55865).

PURPOSE: The study was designed to determine independent prognostic variables in suboptimally debulked advanced ovarian cancer patients entered in the randomised phase III study EORTC 55865. EXPERIMENTAL DESIGN: Retrospectively collected paraffin blocks from 169 patients with stages IIb-IV epithelial ovarian cancer, taken at primary debulking surgery, were analysed. All patients were treated with cyclophosphamide and cisplatin (CP), and followed up for a median of 10 years. Expression of p53, bcl-2, P21, Ki-67 and HER-2 status was assessed by immunohistochemistry (IHC). RESULTS: Expression of p21, a downstream effector of the p53 gene, was found to be a favourable prognostic factor for survival (HR 0.58, CI 0.36-0.94, p=0.025) in addition to FIGO stage (HR 1.54, CI 1.08-2.21, p=or<0.02). For progression free survival (PFS), both p21 (HR 0.52) and Ki-67 (HR 0.6) were significant factors. CONCLUSION: P21 overexpression is a positive prognostic factor for survival and PFS in advanced ovarian carcinoma with residual lesions of more than 1 cm.

Adult↗

Predicting rate of oxygen consumption from heart rate while little penguins work, rest and play.

The relationship between heart rate (f(H)) and rate of oxygen consumption (V(.)O2) was investigated under changing conditions of ambient temperature, digestive state and exercise state in the little penguin (Eudyptula minor). Both f(H) and V(.)O2 were recorded simultaneously from 12 little penguins while they each (a) rested and exercised within their reported thermo-neutral zone (TNZ), (b) rested and exercised below their reported TNZ and (c) digested a meal of sardines within their reported TNZ. Contrary to our expectations, we found that minimum V(.)O2 did not vary between the two temperatures used. Comparison with values from the literature suggests that both minimum V(.)O2 and the extent of the TNZ in this species may vary along a latitudinal gradient. Furthermore, while minimum V(.)O2 was unchanged at the lower temperature, minimum f(H) was significantly higher, suggesting a hitherto undescribed cardiac response to lowered ambient temperature in an avian species. This response was maintained when the penguins exercised within and below their apparent TNZ as f(H) was significantly greater in cold conditions for a given level of V(.)O2. Furthermore, both f(H) and V(.)O2 were slightly but significantly elevated for a given walking speed during exercise at the lower temperature. This suggests that the penguins may have been close to their TNZ and that the measures employed to counteract heat loss while at rest may have been compromised during exercise. There was no significant difference in the relationship between f(H) and V(.)O2 while the penguins were inactive ina post-digestive state or inactive and digesting a meal within their TNZ, though both of these relationships were significantly different from that during exercise. This suggests that while digestion has no effect on the f(H)/V(.)O2 relationship, for little penguins at least, it is of little value in deriving a predictive relationship for application to active free-ranging animals.

Animals↗

Increased vascular endothelial growth factor and pregnancy-associated glycoproteins, but not insulin-like growth factor-I, in maternal blood of cows gestating twin fetuses.

Differences in placental mass and vascularity exist between cows gestating single vs. multiple fetuses. Therefore, the association between fetal number and placental development or function was assessed by comparing concentrations of vascular endothelial growth factor (VEGF), pregnancy-associated glycoproteins (PAG), IGF-I, and progesterone in the maternal blood of cattle selected for twin births and gestating 1 (n = 23) vs. 2 (n = 17) fetuses. Samples of jugular venous blood were collected serially at a mean of 57, 121, 192, and 234 d (range within groups was 20 d) after AI. Plasma concentrations of VEGF, IGF-I, and progesterone were measured by double-antibody RIA, and of PAG by an indirect sandwich ELISA. Concentrations of VEGF and progesterone were greater (P < 0.05) in dams with twin vs. single fetuses. Maternal VEGF concentrations did not differ among collection times, but progesterone concentrations increased (P < 0.01) between d 192 and 234. Conversely, PAG concentrations were low at d 57 and 121 and did not differ between dams carrying singles or twins. However, the subsequent increase (P < 0.01) in PAG was greater in dams with twins, resulting in greater (P < 0.01) PAG concentrations for dams with twins at d 192 and 234 (type of birth x time; P < 0.01). Maternal IGF-I concentrations were unaffected by fetal number. Because corpora lutea persisted for the duration of the evaluation period, maternal progesterone concentrations were likely related to the number of corpora lutea rather than the number of fetuses. It is postulated that the greater PAG and VEGF concentrations in the blood of dams gestating twins are the result of a larger uteroplacental mass, including increased numbers of binucleate cells and increased angiogenesis and vasculogenesis associated with a twin pregnancy. Although PAG and VEGF were elevated in dams gestating twins, variability within and among birth groups limits the use of PAG or VEGF measurements for the diagnosis of twins.

Animals↗

The use of microarrays to define functionally-related genes that are differentially expressed in the cycling pig uterus.

In swine and other livestock, the uterine endometrium exhibits dramatic morphological and secretory changes throughout the oestrous cycle and during pregnancy. Such physiological changes are a reflection of extremely complex interactions between gene products (RNA and protein). The recent development of genomics and proteomics methods, as well as associated bioinformatics tools, has provided the means to begin characterising such interactions. Indeed, the analysis of the transcriptome and proteome of cells and tissues now comprises a new field of study known as 'systems biology'. Currently, the most powerful technique available to the systems biologist is the microarray. These platforms represent oligonucleotide or cDNA fragments spotted in a specified high-density pattern on a solid support. Hybridisation of fluorescently-tagged cDNAs from different tissue sources permits the measurement of thousands of RNAs in parallel. The method permits the identification of genes that are present at different amounts between the two tissues and, more importantly, it permits the identification of groups of genes (clusters) that are expressed in comparable patterns. Results from a recent expression profiling experiment are described. The goal of the profiling experiment was to define genes that are differentially expressed in endometrium during the oestrous cycle. The experiment used an in-house cDNA microarray with > 14,000 distinct cDNAs cloned from reproductive tissues. Total RNAs from cyclic endometrium (Days 0, 3, 6, 10, 12, 14 and 18 post-oestrus) were reverse transcribed into cDNAs, labelled with fluorescent dye and hybridised to the arrays along with cDNAs derived from a reference RNA pool. A total of 4,827 genes were found to differ significantly at some time during the oestrous cycle. Clustering methods were able to define numerous groups of similarly expressed genes. These data will help to define the complex patterns of endometrial genes acting in concert to create the environments required for fertilisation, embryo growth and conceptus development in swine.

Animals↗

Filter sterilization of highly infectious samples to prevent false negative analysis of matrix metalloproteinase activity.

Matrix metalloproteinases (MMPs) are implicated in the immunopathology of numerous infectious diseases. High risk samples such as those generated after infection with Mycobacterium tuberculosis require filter sterilization for safe analysis of MMP concentrations. Here, we report that commercial filter membranes may cause artefacts by binding MMPs. Anopore 0.2 microM membrane filtration reduced MMP-1 concentrations to undetectable levels by zymography and Western blotting. Polypropylene 0.45 microM filtration removed some MMP-1, while Polysulphone, Durapore and Bio-inert 0.2 microM membranes did not remove MMP-1. Anopore filtration also removed all MMP-7 and -9 activity, suggesting that the conserved MMP catalytic domain binds the membrane. This study demonstrates the importance of selecting the appropriate filter in MMP analysis to avoid incorrectly excluding MMP involvement in infection-related immunopathology.

Binding Sites↗

Allometric estimation of metabolic rate from heart rate in penguins.

Studies of the relationship between heart rate (f(H)) and rate of oxygen consumption (V(.) (O(2))), which are then used to predict field metabolic rate, frequently fail to incorporate body mass as a predictive variable. This is a potentially important omission in the study of animals whose body mass fluctuates substantially during their annual cycle. In an attempt further to improve estimates of field metabolic rate from f(H), we re-evaluated data on M(b), f(H) and V(.) (O(2)) from previous studies of macaroni penguins (Eudyptes chrysolophus) and king penguins (Aptenodytes patagonicus) and derived a new relationship to integrate these three quantities. This relationship is at least as accurate and precise as previously determined relationships. We applied this same principle to published data on 11 of the 20 recognised penguin taxa to derive a relationship to predict V(.) (O(2)) from f(H) and M(b) in penguins of any species. This result has interesting implications in terms of reducing the logistical burden in studies of field metabolic rate.

Animals↗

Transcriptional profiling of pig embryogenesis by using a 15-K member unigene set specific for pig reproductive tissues and embryos.

Differential mRNA expression patterns were evaluated between germinal vesicle oocytes (pgvo), four-cell (p4civv), blastocyst (pblivv), and in vitro-produced four-cell (p4civp) and in vitro-produced blastocyst (pblivp) stage embryos to determine key transcripts responsible for early embryonic development in the pig. Five comparisons were made: pgvo to p4civv, p4civv to pblivv, pgvo to pblivv, p4civv to p4civp, and pblivv to pblivp. ANOVA (P < 0.05) was performed with the Benjamini and Hochberg false-discovery-rate multiple correction test on each comparison. A comparison of pgvo to p4civv, p4civv to pblivv, and pgvo to pblivv resulted in 3214, 1989, and 4528 differentially detected cDNAs, respectively. Real-time PCR analysis on seven transcripts showed an identical pattern of changes in expression as observed on the microarrays, while one transcript deviated at a single cell stage. There were 1409 and 1696 differentially detected cDNAs between the in vitro- and in vivo-produced embryos at the four-cell and blastocyst stages, respectively, without the Benjamini and Hochberg false-discovery-rate multiple correction test. Real-time polymerase chain reaction (PCR) analysis on four genes at the four-cell stage showed an identical pattern of gene expression as found on the microarrays. Real-time PCR analysis on four of five genes at the blastocyst stage showed an identical pattern of gene expression as found on the microarrays. Thus, only 1 of the 39 comparisons of the pattern of gene expression exhibited a major deviation between the microarray and the real-time PCR. These results illustrate the complex mechanisms involved in pig early embryonic development.

Animals↗

Use of chromogenic medium in the isolation of yeasts from clinical specimens.

Over a 1 year period 3296 specimens submitted for fungal culture were plated onto routine mycological media (RM) and CHROMagar Candida (CaC) to evaluate the capability of CaC to improve on RM. With RM, cultures producing single yeast isolates were identified from 802 specimens. CaC produced similar results, with 76 % agreement. Of 761 specimens that yielded a single Candida species by RM, 615 (81 %) produced one or more yeast isolates using CaC. Of concern, 132 negative CaC cultures corresponded to specimens that yielded C. albicans alone on RM. When yeasts were recovered, CaC correctly identified 98 % of C. albicans, 93 % of Candida tropicalis, 96 % of Candida glabrata and 100 % of Candida krusei based on typical colours. CaC did potentially improve on RM by detecting yeasts in 91 specimens that yielded none by routine methods. CaC was noted to recover more yeast isolates than RM when mixed cultures were detected. Overall, the role of CaC in improving RM appears limited.

Blood↗

Do seasonal changes in metabolic rate facilitate changes in diving behaviour?

Macaroni penguins were implanted with data loggers to record heart rate (fH), abdominal temperature (Tab) and diving depth during their pre-moult trip (summer) and winter migration. The penguins showed substantial differences in diving behaviour between the seasons. During winter, mean and maximum dive duration and dive depth were significantly greater than during summer, but the proportion of dives within the calculated aerobic dive limit (cADL) did not change. Rates of oxygen consumption were estimated from fH. As winter progressed, the rate of oxygen consumption during dive cycles (sVO2DC)) declined significantly and mirrored the pattern of increase in maximum duration and depth. The decline in sVO2DC) was matched by a decline in minimum rate of oxygen consumption (sVO2min)). When sVO2min) was subtracted from sVO2DC), the net cost of diving was unchanged between summer and winter. We suggest that the increased diving capacity demonstrated during the winter was facilitated by the decrease in sVO2min). Abdominal temperature declined during winter but this was not sufficient to explain the decline in sVO2min). A simple model of the interactions between sVO2min), thermal conductance and water temperature shows how a change in the distribution of fat stores and therefore a change in insulation and/or a difference in foraging location during winter could account for the observed reduction in sVO2min) and hence sVO2DC).

Animals↗

A randomised controlled trial of oral zinc on the immune response to tuberculosis in HIV-infected patients.

SETTING: The National HIV Unit, Singapore. OBJECTIVE: To test whether zinc supplementation improves the immune response to tuberculosis in HIV-positive patients. DESIGN: A double-blind, randomised, placebo-controlled trial of 28 days of oral zinc sulphate (50 mg of elemental zinc) or placebo in stable adult HIV-positive patients receiving antiretroviral therapy with a CD4 count <200 cells/microl. METHODS: IFN-gamma response to mycobacterial antigen stimulation, CD4/8 cell count, lymphocyte subsets, T-cell receptor excision circle (TREC) levels and viral load were measured at baseline and day 28. RESULTS: Thirty-two patients received zinc and 34 placebo. There was no significant change in the IFN-gamma response to human PPD stimulation in the zinc or placebo groups (placebo baseline: 0.42 +/- 1.03, day 28: 0.84 +/- 1.21 IU/ml, zinc baseline: 1.26 +/- 2.41, day 28: 1.39 +/- 1.88 IU/ml, P = 0.31 between groups), nor any of the other mycobacterial antigens tested. There were no changes in absolute CD4/8 cell levels or other lymphocyte subsets, TREC or viral load. Baseline zinc levels were normal in 62/66 (93.9%) patients. CONCLUSIONS: We found no evidence for recommending pharmacological supplementation with oral zinc in HIV-positive patients without zinc deficiency.

Administration, Oral↗

Light and electron microscope immunocytochemical studies of the distribution of pregnancy associated glycoproteins (PAGs) throughout pregnancy in the cow: possible functional implications.

Pregnancy associated glycoproteins (PAGs) comprise a large group of placental antigens of the aspartic proteinase family. Phylogenetic analysis indicated that the PAGs form two distinct groups, one of ancient origin and one produced by a more recent series of gene duplications. This paper summarises the molecular biological and biochemical studies which have been used to purify and raise antibodies against specific PAGs and groups of related PAGs and their use in light and electron microscope immunocytochemistry to demonstrate that the ancient PAG group has a similar distribution at the placental fetomaternal interface (microvillar junction, MVJ) in cows and pigs. This localization suggests either a possible role in binding the surfaces together and/or in establishment of an immunological barrier. The more recently evolved PAG group, absent in the pig, exhibited no significant localization to the MVJ but was restricted to the trophoblast binucleate cell (BNC) granules in the cow. Furthermore, these PAGs bind to newly formed maternal uterine connective tissue to which they are delivered by BNC migration and fusion with uterine epithelial cells. At this location in the developing maternal villi of the placentomes, they are ideally positioned to manipulate the maternal immune system to facilitate a successful pregnancy.

Animals↗