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Biomedical subjects

J Lamb

Publications and source records attributed to J Lamb.

At least 19 recordsLinked to original sources

Acid-adaptation does not increase the resistance of Listeria monocytogenes to irradiation in a seafood salad.

Stress adaptation of microbial cells enables the cells to survive better when they are subsequently exposed to other types of stresses. In the food industry, pathogens are commonly stressed during food processing and this is a concern where pathogens such as Listeria monocytogenes are involved. Research was conducted to determine if acid adaptation of L. monocytogenes provides resistance to ionizing irradiation. Three different strains of L. monocytogenes were acid-adapted using three different acids (acetic, citric, lactic) in Tryptic Soy Broth, at a pH 5.5 for 1 h, 4 h, or continuous acid exposure. The acid-adapted L. monocytogenes were then exposed to a low level of gamma irradiation (0.59-0.72 kGy) along with a non-acid adapted L. monocytogenes control. In a test tube study, the 1-h acetic acid-adapted L. monocytogenes strains showed the greatest difference from the control, a reduced kill of 1.1 log CFU/g but this difference was not significant by ANOVA (p=0.054). The reduction achieved after 4 h and continuous acid exposure also did not significantly differ from the control. To determine whether acid adaptation affected radiation resistance within a food product, a refrigerated storage shelf-life study was completed. Acetic acid was used to acid adapt a three-strain cocktail of L. monocytogenes for a period of 1 h. The organisms were then inoculated into a seafood salad (pH 5.15) and subsequently exposed to low dose gamma irradiation (0.7 to 4.5 kGy). L. monocytogenes was reduced or eliminated by irradiation regardless of acid adaptation; no increased resistance was observed.

Acetic Acid↗

The role of the spleen in laparoscopy-associated inflammatory response.

BACKGROUND: Carbon dioxide (CO(2)) pneumoperitoneum alters the inflammatory response in animal models of sepsis. The spleen is a key organ in inflammation and its removal was predicted to modify this effect. METHODS: The acute phase inflammatory response to lipopolysaccharide (LPS) challenge in male rats was examined for the effects of splenectomy (spx) and the technique of removal (open or laparpscopic). A series of experiments compared LPS-only controls with LPS injection 2 or 9 days following open spx, lap CO2 spx, open sham, or lap CO2 sham. The method of splenectomy was studied by randomization to control, open spx, lap CO2 spx, lap helium (He) spx, or lap air spx with LPS challenge on postoperative day 2. Serum levels of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and, interleutin (IL) 10 were collected at multiple time points, assayed by commercial enzyme-linked immunosorbent assay, analyzed by analysis of variance. RESULTS: Levels of TNF-alpha at 1.5 were significantly lower following open sham than following lap sham (p < 0.05). Splenectomy drastically reduced INF-gamma and TNF-alpha levels compared to controls (p < 0.05) on postoperative day 2. No method of spx preserved TNF-alpha or INF-gamma responses. Recovery of TNF-alpha response on day 9 was delayed in the spx groups. CONCLUSIONS: Splenectomy dramatically reduces TNF-alpha and INF-gamma responses to LPS challenge, although by different mechanisms. Pneumoperitoneum-mediated modulation of the septic inflammatory response is partially dependent on the spleen.

Acute-Phase Reaction↗

Clinical and cost-effectiveness of autologous chondrocyte implantation for cartilage defects in knee joints: systematic review and economic evaluation.

OBJECTIVE: To support a review of the guidance issued by the National Institute for Health and Clinical Excellence (NICE) in December 2000 by examining the current clinical and cost-effectiveness evidence on autologous cartilage transplantation. DATA SOURCES: Electronic databases. REVIEW METHODS: Evidence on clinical effectiveness was obtained from randomised trials, supplemented by data from selected observational studies for longer term results, and for the natural history of chondral lesions. Because of a lack of long-term results on outcomes such as later osteoarthritis and knee replacement, only illustrative modelling was done, using a range of assumptions that seemed reasonable, but were not evidence based. RESULTS: Four randomised controlled trials were included, as well as observational data from case series. The trials studied a total of 266 patients and the observational studies up to 101 patients. Two studies compared autologous chondrocyte implantation (ACI) with mosaicplasty, the third compared ACI with microfracture, and the fourth compared matrix-guided ACI (MACI) with microfracture. Follow-up was 1 year in one study, and up to 3 years in the remaining three studies. The first trial of ACI versus mosaicplasty found that ACI gave better results than mosaicplasty at 1 year. Overall, 88% had excellent or good results with ACI versus 69% with mosaicplasty. About half of the biopsies after ACI showed hyaline cartilage. The second trial of ACI versus mosaicplasty found little difference in clinical outcomes at 2 years. Disappointingly, biopsies from the ACI group showed fibrocartilage rather than hyaline cartilage. The trial of ACI versus microfracture also found only small differences in outcomes at 2 years. Finally, the trial of MACI versus microfracture contained insufficient long-term results at present, but the study does show the feasibility of doing ACI by the MACI technique. It also suggested that after ACI, it takes 2 years for full-thickness cartilage to be produced. Reliable costs per quality-adjusted life-year (QALY) could not be calculated owing to the absence of necessary data. Simple short-term modelling suggests that the quality of life gain from ACI versus microfracture would have to be between 70 and 100% greater over 2 years for it to be more cost-effective within the 20,000--30,000 pounds sterling per QALY cost-effectiveness thresholds. However, if the quality of life gains could be maintained for a decade, increments relative to microfracture would only have to be 10--20% greater to justify additional treatment costs within the cost-effectiveness band indicated above. Follow-up from the trials so far has only been up to 2 years, with longer term outcomes being uncertain. CONCLUSIONS: There is insufficient evidence at present to say that ACI is cost-effective compared with microfracture or mosaicplasty. Longer term outcomes are required. Economic modelling using some assumptions about long-term outcomes that seem reasonable suggests that ACI would be cost-effective because it is more likely to produce hyaline cartilage, which is more likely to be durable and to prevent osteoarthritis in the longer term (e.g. 20 years). Further research is needed into earlier methods of predicting long-term results. Basic science research is also needed into factors that influence stem cells to become chondrocytes and to produce high-quality cartilage, as it may be possible to have more patients developing hyaline cartilage after microfracture. Study is also needed into cost-effective methods of rehabilitation and the effect of early mobilisation on cartilage growth.

Cartilage, Articular↗

Initial chest tube management after pulmonary resection.

Tube thoracostomy management with suction or water seal after anatomical pulmonary resection remains somewhat controversial. Initial chest tube management may influence the duration of pleural fluid drainage, duration of tube thoracostomy, and/or hospital length of stay following pulmonary resection. We hypothesized that initial chest tube management with water seal decreases time for chest tube removal and decreases time of hospital stay. A retrospective chart review was performed on 109 consecutive patients who underwent lobectomy or segmentectomy in Western Pennsylvania Hospital between December 1999 and December 2003. Comparison was made between chest tube management of water seal or suction in patients with and without air leak. Of the 109 patients, 78 (72%) had no air leak at the completion of surgery, and 31 (28%) had air leak. In the group without air leak (n = 78), water seal was used in 32 (41%) patients and suction in 46 (59%). In patients placed to water seal initially after surgery (n = 32), removal of chest tubes was on postoperative day (POD) 3.19 +/- 0.24 and hospital discharge was on POD 5.13 +/- 0.61. In patients placed to suction initially (n = 46), chest tubes were removed on POD 4.52 +/- 0.40. Hospital discharge was on POD 6.74 +/- 0.5. Both duration of chest tube (P < 0.007) and length of hospital stay (P < 0.04) were significantly lower in the water seal group. In the air leak group (n = 31), 7 (23%) patients were managed with water seal and 24 (77%) patients with suction. Both duration of chest tube (P = 0.001) and length of hospital stay (P < 0.05) were significantly lower in the water seal group. In patients without air leak, chest tubes should be managed with water seal following anatomical pulmonary resection, resulting in significantly shorter chest tube duration and hospital length of stay.

Adult↗

The effect of timing of pneumoperitoneum on the inflammatory response.

BACKGROUND: We examined the effects of an identical period of pneumoperitoneum applied at three different time points after lipopolysaccharide (LPS) challenge. Two different insufflation gases were also compared. METHODS: Male rats (n = 70) were injected intravenously with 1 mg/kg of LPS (time 0). The time relationship between a 1.5-h period of insufflation and initial LPS stimulation was the experimental variable. All rats were killed 6 h after injection. CO2 and helium insufflation were investigated. Ten control rats received LPS only. Serum interleukin-6 (IL-6) levels were determined by enzyme-linked immunosorbent assay (ELISA). Hepatic expression of alpha2-macroglobulin, beta-fibrinogen, and metallothionein were measured by Northern blot analysis. Statistical analysis was performed using one-way analysis of variance (ANOVA). RESULTS: Expression of alpha2-macroglobulin mRNA was lower in CO2 groups compared to the control group (p < 0.05 at time 120 and 270). beta-Fibrinogen message was diminished in CO2 0 and 120 groups compared to control. Serum levels of IL-6 and expression of metallothionein mRNA did not show significant differences between groups. CONCLUSIONS: These findings suggest that CO2 pneumoperitoneum downregulates the inflammatory response to LPS challenge. Start time of CO2 insufflation does not appear to alter hepatic expression of acute phase genes. The mechanism of alpha2-macroglobulin downregulation does not appear to be due to IL-6.

Animals↗

An integrative genomics approach to the reconstruction of gene networks in segregating populations.

The reconstruction of genetic networks in mammalian systems is one of the primary goals in biological research, especially as such reconstructions relate to elucidating not only common, polygenic human diseases, but living systems more generally. Here we propose a novel gene network reconstruction algorithm, derived from classic Bayesian network methods, that utilizes naturally occurring genetic variations as a source of perturbations to elucidate the network. This algorithm incorporates relative transcript abundance and genotypic data from segregating populations by employing a generalized scoring function of maximum likelihood commonly used in Bayesian network reconstruction problems. The utility of this novel algorithm is demonstrated via application to liver gene expression data from a segregating mouse population. We demonstrate that the network derived from these data using our novel network reconstruction algorithm is able to capture causal associations between genes that result in increased predictive power, compared to more classically reconstructed networks derived from the same data.

11-beta-Hydroxysteroid Dehydrogenases↗

Combined monitoring of motor and somatosensory evoked potentials in orthopaedic spinal surgery.

OBJECTIVES: To demonstrate possible advantages of combined (motor and sensory) versus single modality (either motor or sensory) intra-operative spinal cord monitoring and to investigate risk factors for post-operative neurological sequelae. METHODS: Recordings of lower limb motor evoked potentials (MEPs) to multi-pulse transcranial electrical stimulation (TES), and tibial nerve somatosensory evoked potentials (SEPs), were attempted during 126 operations in 97 patients (79 with spinal deformity and 18 with miscellaneous spinal disorders). RESULTS: Combined motor and sensory monitoring was successfully achieved in 104 of 126 (82%) operations. No response to either modality could be recorded in two patients with Friedreich' s ataxia. In 18 patients monitoring was possible in only one modality: SEPs could not be recorded in two patients and MEPs in 16. Significant intra-operative EP changes occurred in one or both modalities in 16 patients; in association with instrumentation in 10 cases, and with systemic changes in 6. After appropriate remedial measures, SEPs recovered either fully or partially in 8/8 patients and MEPs in only 67% (10/15 patients). New deficits were present post-operatively in 6 of the 16 patients with abnormal intra-operative EPs. Normal MEPs at the end of the operation correctly predicted the absence of new motor deficits in all cases. SEPs either remained unchanged or recovered fully after remedial measures in 3 patients with new post-operative motor deficits. Neurological complications were more frequent in patients with miscellaneous spinal disorders and/or pre-existing neurological deficits. No complications occurred in patients with idiopathic scoliosis. CONCLUSIONS: Combined SEPs and multi-pulse TES-MEPs provide a safe, reliable and sensitive method of monitoring spinal cord function in orthopaedic surgery. This method is superior to single modality techniques, both for increasing the number of patients in whom satisfactory monitoring of spinal cord function can be achieved and, for improving the sensitivity and predictivity of monitoring. Combined SEP/MEP methods may enhance the impact of neuromonitoring on the intra-operative management of the patient and favourably influence neurological outcome.

Action Potentials↗

Estimation and management of genetic diversity in small populations of plains zebra (Equus quagga) in KwaZulu-Natal, South Africa.

Plains zebras (Equus quagga antiquorum) occur in few large, but many small, isolated populations in KwaZulu-Natal. Problems identified in small populations include reduced striping patterns on hind quarters, smaller size, elevated mortality rates and high number of still-births. Inbreeding may be implicated. Population viability analysis (PVA) was conducted with a computer model (VORTEX), and DNA and allozyme analyses were conducted to test the findings of the model. Using standard methods, DNA (PCR-RAPD) and allozyme diversity was assessed in blood samples from 72 plains zebra from four KwaZulu-Natal Nature Conservation Services (KZN-NCS) protected areas: Umfolozi Game Reserve (UGR), Albert Falls (AFNR), Vernon Crookes (VCNR) and Harold Johnson (HJNR) Nature Reserves. Populations of the latter three, small-sized (9-110 individuals) populations were seeded from the same source population (UGR: current population of 2000) during the past 25 years. Information from PCR-RAPD and allozyme analyses were compared with each other as well as to that predicted by population genetic modelling (using VORTEX). Allozyme heterozygosities were consistently high in all populations (12.1-12.9%), with no observable losses associated with reduced population size. On the other hand, percentage loss of polymorphism (20-39%) calculated from the PCR-RAPD study appeared to be positively correlated with the loss of heterozygosity predicted by population viability analysis (PVA), and negatively correlated with population size. On the basis of the above results, a policy of translocation was advocated for small, intensely managed populations of zebras, whereby a harem should be translocated every five years for a population size of nine (HJNR), while for a population size of 110 (VCNR) translocations should take place every 15 years if heterozygosity is to be maintained at more than 90% within each population over 100 years.

Journal Article↗

Adolescent research recruitment issues and strategies: application in a rural school setting.

This report discusses adolescent research recruitment strategies in general and specifically, those developed by a research team for a rural school setting. Recruitment of adolescent subjects involves the consideration of several complex issues. These issues include adolescent development, ethical and legal guidelines, and access to school settings. Specific strategies are discussed regarding access and culture of the rural setting, collaboration, and recruitment procedures in relation to their use to recruit adolescent subjects from rural schools.

Adolescent↗

Detection of Kaposi's sarcoma-associated herpesvirus in oral and genital secretions of Zimbabwean women.

Kaposi's sarcoma-associated herpesvirus (KSHV) in oral and genital secretions of women may be involved in horizontal and vertical transmission in endemic regions. Nested polymerase chain reaction assays were used to detect KSHV DNA sequences in one-third of oral, vaginal, and cervical specimens and in 42% of peripheral blood mononuclear cell (PBMC) specimens collected from 41 women infected with human immunodeficiency virus type 1 who had Kaposi's sarcoma (KS). KSHV DNA was not detected in specimens from 100 women without KS, 9 of whom were seropositive for KSHV. A positive association was observed between KSHV DNA detection in oral and genital mucosa, neither of which was associated with KSHV DNA detection in PBMC. These data suggest that KSHV replicates in preferred anatomic sites at levels independent of PBMC viremia. Detection of genital-tract KSHV only among relatively immunosuppressed women may provide an explanation for infrequent perinatal transmission of KSHV.

AIDS-Related Opportunistic Infections↗

Single amino acid (arginine) deprivation induces G1 arrest associated with inhibition of cdk4 expression in cultured human diploid fibroblasts.

Withdrawal of a single amino acid (arginine) from freely cycling early passage primary human fibroblasts caused a halt to proliferation, characterized by an accumulation of cells in the G1 phase of the cell cycle. This arrest was accompanied by the suppression of cyclin D1- and cyclin E-associated kinase activities and the appearance of hypophosphorylated retinoblastoma protein. Arginine-deprived cells remained viable for in excess of 4 days and could be made to synchronously reenter the cell cycle by restoration of the amino acid, with kinetics characteristic of exit from a quiescent state. Stimulation of cells arrested by serum withdrawal did not result in S-phase entry when arginine was omitted from the culture medium. Although cyclin D1 accumulated on normal schedule, cdk4, which increased following restimulation in amino acid-replete medium, was not induced when arginine was absent. These results suggest that arginine deprivation-in common with other "suboptimal" conditions-inhibits the passage of normal human cells through the restriction point and implicate cdk4 as the key regulatory element in amino acid-sensitive cell cycle control.

Amino Acid Substitution↗

Pathology characteristics that optimize outcome prediction of a breast screening trial.

The ability of pathology characteristics to predict outcome was tested with the 1029 cancers accumulated in the Edinburgh Randomized Trial of breast screening after 14 years follow-up. The majority (55.7%) were in the screening arm, which also had more operable cases (81.3% vs 62.2%); the reduction in the proportion of inoperable breast cancers in a UK female population invited to mammographic screening is a notable effect of the trial. In the 691 operable invasive cases the size, histological type, grade, node status and node number group individually showed highly significant (P<0.001) association with survival. In multivariate analysis the Nottingham Prognostic Index (NPI) derived from these features showed highly significant association with survival (P<0.001). However, when first adjusted for NPI, combined addition of pathological size in 6 categories and histological type as special or not had an independent association with survival that was statistically firmly based (P<0.001). For operable breast cancer the gains are in smaller sizes, better histological features, and higher proportion node negative. The weighting factors applied to pathology indicators of survival in the NPI are not optimal for a population included in a trial of screening. In particular, a linear trend of the index with pathological size is not appropriate. Inclusion of histological type as special or not improves the index further.

Analysis of Variance↗

Single amino acid (arginine) deprivation: rapid and selective death of cultured transformed and malignant cells.

The effects of arginine deprivation (-Arg) has been examined in 26 cell lines. Less than 10% of those with transformed or malignant phenotype survived for > 5 days, and many died more rapidly, notably leukaemic cells. Bivariate flow cytometry confirmed that vulnerable cell lines failed to move out of cell cycle into a quiescent state (G0), but reinitiated DNA synthesis. Many cells remained in S-phase, and/or had difficulty progressing through to G2 and M. Two tumour lines proved relatively 'resistant', A549 and MCF7. Although considerable cell loss occurred initially, both lines showed a 'cell cycle freeze', in which cells survived for > 10 days. These cells recovered their proliferative activity in +Arg medium, but behaved in the same manner to a second -Arg episode as they did to the first episode. In contrast, normal cells entered G0 and survived in -Arg medium for several weeks, with the majority of cells recovering with predictable kinetics in +Arg medium. In general, cells from a wide range of tumours and established lines die quickly in vitro following -Arg treatment, because of defective cell cycle checkpoint stringency, the efficacy of the treatment being most clearly demonstrated in co-cultures in which only the normal cells survived. The findings demonstrate a potentially simple, effective and non-genotoxic strategy for the treatment of a wide range of cancers.

Animals↗

Regulation of the functional interaction between cyclin D1 and the estrogen receptor.

We report that the functional interaction between cyclin D1 and the estrogen receptor (ER) is regulated by a signal transduction pathway involving the second messenger, cyclic AMP (cAMP). The cell-permeable cAMP analogue 8-bromo-cAMP caused a concentration-dependent enhancement of cyclin D1-ER complex formation, as judged both by coimmunoprecipitation and mammalian two-hybrid analysis. This effect was paralleled by increases in ligand-independent ER-mediated transcription from an estrogen response element containing reporter construct. These effects of 8-bromo-cAMP were antagonized by a specific protein kinase A (PKA) inhibitor, indicating that the signaling pathway involved was PKA dependent. Further, we show that culture of MCF-7 cells on a cellular substratum of murine preadipocytes also enhanced the functional interaction between cyclin D1 and ER in a PKA-dependent manner. These findings demonstrate a collaboration between cAMP signaling and cyclin D1 in the ligand-independent activation of ER-mediated transcription in mammary epithelial cells and show that the functional associations of cyclin D1 are regulated as a function of cellular context.

8-Bromo Cyclic Adenosine Monophosphate↗

Single amino acid (arginine) restriction: growth and death of cultured HeLa and human diploid fibroblasts.

Requirements for arginine are different from leucine for the growth of HeLa cells in monolayer and suspension culture. Cells grow increasingly more slowly as arginine levels fall below millimolar. Most cells died at 10(-5) M in static cultures, but could be sustained in perfused cultures, but at 10(-6) M neither perfusion nor increased volume in static cultures compensated. Cell died within 3-4 days in 10(-6) M in the same manner as those in complete arginine deprivation, i.e. considerably faster than with leucine deprivation. Arginine restriction produced by arginase or arginine decarboxylase addition to culture medium gave similar results. Citrulline substituted for arginine, but ornithine and polyamines did not. Arginine was depleted 3-4 times faster from the medium than other amino acids, <5% being consumed in protein synthesis, and arginine released by protein turnover was less efficiently reutilised than leucine. Deprivation reduced protein and DNA syntheses, greatly extended S-phase and protracted the cell cycle in HeLa cells for more than leucine deprivation. The inability of the cells avoid reinitiation of S-phase resulted in their proliferative impetus driving them into an late cycle (premitotic) death. In contrast, normal human diploid fibroblasts reached quiescence with little delay and survived for >11 days. Arginine deprivation is discussed as a selectively means of tumour cell destruction.

Arginine↗

Sustained in vivo regression of Dunning H rat prostate cancers treated with combinations of androgen ablation and Trk tyrosine kinase inhibitors, CEP-751 (KT-6587) or CEP-701 (KT-5555).

The indolocarbazole analogue CEP-751 is a potent and selective tyrosine kinase inhibitor of the neurotrophin-specific trk receptors that has demonstrated antitumor activity in nine different models of prostate cancer growth in vivo. In the slow-growing, androgen-sensitive Dunning H prostate cancers, which express trk receptors, CEP-751 induced transient regressions independent of effects on cell cycle. Because androgen ablation is the most commonly used treatment for prostate cancer, we examined whether the combination treatment of CEP-751 with castration would lead to better antitumor efficacy than either treatment alone. For a 60-day period, H tumor-bearing rats received treatment with either castration, CEP-751 (10 mg/kg once a day s.c. for 5 days every 2 weeks), a combination of both, or vehicle. Castration caused tumor regression, followed by tumor regrowth in 4-6 weeks, whereas intermittent CEP-751 treatments resulted in tumor regressions during each treatment, which were followed by a period of regrowth between intermittent drug treatment cycles. Overall, both monotherapies significantly inhibited tumor growth compared with the vehicle-treated control group. However, the combination of castration and concomitant CEP-751 produced the most dramatic results: sigificantly greater tumor regression than either therapy alone, with no signs of regrowth. A related experiment using an orally administered CEP-751 analogue (CEP-701), as the trk inhibitor, and a gonadotrophin-releasing hormone agonist, Leuprolide, to induce androgen ablation demonstrated similar results, indicating that these effects could be generalized to other forms of androgen ablation and other trk inhibitors within this class. In addition, when CEP-701 was given sequentially to rats bearing H tumors, which were progressing in the presence of continuous androgen ablation induced by Leuprolide, regression of the androgen-independent tumors occurred. In summary, these data demonstrate that CEP-751 or CEP-701, when combined with surgically or chemically induced androgen ablation, offer better antitumor efficacy than either monotherapy and suggest that each therapy produces prostate cancer cell death through complementary mechanisms.

Adenocarcinoma↗

Screening and predicting adolescent depressive symptoms in rural settings.

Adolescent depression with related suicidal behavior is a serious health problem. This article describes depressive symptoms, reported life events, and demographic risk factors in 846 rural adolescents. The results indicate that self-reported depressive symptoms in rural adolescents is significantly (p < .05) related to gender, death in the family, and the perceived positive and negative impact of life events, as well as the specific events of losing a close friend, an increase in number of arguments with parents, trouble with classmates, and trouble with police. The type of school program and trouble with siblings was marginally significant (p < .10). These demographic factors and life events may assist health-care professionals in identifying adolescents at risk for depressive symptoms.

Adolescent↗