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1.
Sci Rep ; 12(1): 11146, 2022 07 01.
Article in English | MEDLINE | ID: mdl-35778472

ABSTRACT

This retrospective study of prospectively collected data aimed to identify unique pain and disability trajectories in patients following lumbar discectomy surgery. Patients of this study population presented chiefly with lumbar radiculopathy and underwent discectomy surgery from thirteen sites enrolled in the CSORN registry. Outcome variables of interest included numeric rating scales for leg/back pain and modified Oswestry disability index scores at baseline, 3, 12, and 24 months post-operatively. Latent class growth analysis was used to identify distinct courses in each outcome. Data from 524 patients revealed three unique trajectories for leg pain (excellent = 18.4%, good = 55.4%, poor = 26.3%), disability (excellent = 59.7%, fair = 35.6%, poor = 4.7%) and back pain (excellent = 13.0%, good = 56.4%, poor = 30.6%). Construct validity was supported by statistically significant differences in the proportions of patients attaining the criteria for minimal important change (MIC; 30%) or clinical success in disability (50% or Oswestry score ≤ 22) (p < 0.001). The variable proportions of patients belonging to poor outcome trajectories shows a disconnect between improved disability and persistence of pain. It will be beneficial to incorporate this information into the realm of patient expectation setting in concert with future findings of potential factors predictive of subgroup membership.


Subject(s)
Radiculopathy , Diskectomy , Humans , Pain , Postoperative Period , Radiculopathy/surgery , Retrospective Studies
2.
Eur Cell Mater ; 36: 200-217, 2018 10 25.
Article in English | MEDLINE | ID: mdl-30370912

ABSTRACT

Numerous studies show promise for cell-based tissue engineering strategies aiming to repair painful intervertebral disc (IVD) degeneration. However, clinical translation to human IVD repair is slow. In the present study, the regenerative potential of an autologous nucleus pulposus (NP)-cell-seeded thermoresponsive hyaluronic acid hydrogel in human lumbar IVDs was assessed under physiological conditions. First, agarose-encased in vitro constructs were developed, showing greater than 90 % NP cell viability and high proteoglycan deposition within HA-pNIPAM hydrogels following 3 weeks of dynamic loading. Second, a bovine-induced IVD degeneration model was used to optimise and validate T1ρ magnetic resonance imaging (MRI) for detection of changes in proteoglycan content in isolated intact IVDs. Finally, isolated intact human lumbar IVDs were pre-scanned using the established MRI sequence. Then, IVDs were injected with HA-pNIPAM hydrogel alone or autologous NP-cell-seeded. Next, the treated IVDs were cultured under cyclic dynamic loading for 5 weeks. Post-treatment T1ρ values were significantly higher as compared to pre-treatment scans within the same IVD and region of interest. Histological evaluation of treated human IVDs showed that the implanted hydrogel alone accumulated proteoglycans, while those that contained NP cells also displayed neo-matrix-surrounded cells within the gel. The study indicated a clinical potential for repairing early degenerative human IVDs using autologous cells/hydrogel suspensions. This unique IVD culture set-up, combined with the long-term physiological culture of intact human IVDs, allowed for a more clinically relevant evaluation of human tissue repair and regeneration, which otherwise could not be replicated using the available in vitro and in vivo models.


Subject(s)
Hyaluronic Acid/chemistry , Hydrogels/chemistry , Nucleus Pulposus/transplantation , Organ Culture Techniques , Regeneration , Temperature , Acrylic Resins/chemistry , Animals , Bioreactors , Cattle , Collagen Type I/metabolism , Collagen Type II/metabolism , Compressive Strength , Elastic Modulus , Female , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Nucleus Pulposus/diagnostic imaging , Proteoglycans/metabolism , Transplantation, Autologous , Wound Healing
3.
Sci Total Environ ; 639: 1349-1359, 2018 Oct 15.
Article in English | MEDLINE | ID: mdl-29929300

ABSTRACT

Ecological and human health impairments related to excess nitrogen (N) in streams and rivers remain widespread in the United States (U.S.) despite recent efforts to reduce N pollution. Many studies have quantified the relationship between N loads to streams in terms of N mass and N inputs to watersheds; however, N concentrations, rather than loads, are more closely related to impacts on human health and aquatic life. Additionally, concentrations, rather than loads, trigger regulatory responses. In this study, we examined how N concentrations are related to N inputs to watersheds (atmospheric deposition, synthetic fertilizer, manure applied to agricultural land, cultivated biological N fixation, and point sources), land cover characteristics, and stream network characteristics, including stream size and the extent of lakes and reservoirs. N concentration data were collected across the conterminous U.S. during the U.S. Environmental Protection Agency's 2008-09 National Rivers and Streams Assessment (n = 1966). Median watershed N inputs were 15.7 kg N ha-1 yr-1. Atmospheric deposition accounted for over half the N inputs in 49% of watersheds, but watersheds with the highest N input rates were dominated by agriculture-related sources. Total N input to watersheds explained 42% and 38% of the variability in total N and dissolved inorganic N concentrations, respectively. Land cover characteristics were also important predictors, with wetland cover muting the effect of agricultural N inputs on N concentrations and riparian disturbance exacerbating it. In contrast, stream variables showed little correlation with N concentrations. This suggests that terrestrial factors that can be managed, such as agricultural N use practices and wetland or riparian areas, control the spatial variability in stream N concentrations across the conterminous U.S.

4.
Eur Spine J ; 27(4): 835-840, 2018 04.
Article in English | MEDLINE | ID: mdl-28012079

ABSTRACT

PURPOSE: To determine the significance of each parameter of the revised Tokuhashi score and identify which is associated with survival. BACKGROUND: Spinal metastases are common and can be a challenging medical issue. Treatment options depend on patients' prognosis. Many scoring systems in the literature help estimate prognosis, such as the Tokuhashi, revised Tokuhashi, and Tomita scoring systems. METHODS: A retrospective review of all patients from 2003 to 2012 treated for spinal metastases in one center was conducted. Imaging, pathology, and charts were reviewed to determine the modified Tokuhashi scores. Scores were then compared to the actual documented survival. Univariate and multiple regression analyses were used to assess the importance of each individual parameter and survival time. Linear regression was used to determine the relationship between the Tokuhashi score and weighted Tokuhashi score with survival time. RESULTS: A total of 126 patients were reviewed. All parameters in the revised Tokuhashi score were significantly associated with survival time except for primary site using univariate analysis. Only the number of spinal metastases and metastasis to major organs showed statistical significance when multiple variable analysis was used. CONCLUSION: A number of spinal metastases and metastasis to major organs were the most important predictors of actual survival. Modification to the score based on population characteristics would help better identify patients with spinal metastases that can benefit from surgery.


Subject(s)
Severity of Illness Index , Spinal Neoplasms/diagnosis , Spine/pathology , Adult , Aged , Female , Humans , Linear Models , Male , Middle Aged , Prognosis , Retrospective Studies , Spinal Neoplasms/mortality , Spinal Neoplasms/secondary , Survival Rate
5.
Eur Cell Mater ; 33: 240-251, 2017 03 24.
Article in English | MEDLINE | ID: mdl-28345732

ABSTRACT

Autologous NP cell implantation is a potential therapeutic avenue for intervertebral disc (IVD) degeneration. However, monolayer expansion of cells isolated from surgical samples may negatively impact matrix production by way of dedifferentiation. Previously, we have used a continuous expansion culture system to successfully preserve a chondrocyte phenotype. In this work, we hypothesised that continuous expansion culture could also preserve nucleus pulposus (NP) phenotype. We confirmed that serial passaging drove NP dedifferentiation by significantly decreasing collagen type II, aggrecan and chondroadherin (CHAD) gene expression, compared to freshly isolated cells. Proliferation, gene expression profile and matrix production in both culture conditions were compared using primary bovine NP cells. Both standard culture and continuous culture produced clinically relevant cell populations. However, continuous culture cells maintained significantly higher collagen type II, aggrecan and CHAD transcript expression levels. Also, continuous expansion cells generated greater amounts of proteoglycan, collagen type II and aggrecan protein deposition in pellet cultures. To our surprise, continuous expansion of human intervertebral disc cells - isolated from acute herniation tissue - produced less collagen type II, aggrecan and CHAD genes and proteins, compared to standard culture. Also, continuous culture of cells isolated from young non-degenerate tissue did not preserve gene and protein expression, compared to standard culture. These data indicated that primary bovine and human NP cells responded differently to continuous culture, where the positive effects observed for bovine cells did not translate to human cells. Therefore, caution must be exercised when choosing animal models and cell sources for pre-clinical studies.


Subject(s)
Nucleus Pulposus/cytology , Tissue Engineering/methods , Wound Healing , Adolescent , Adult , Animals , Cattle , Cell Differentiation , Cell Proliferation , Cell Separation , Cells, Cultured , Extracellular Matrix/genetics , Extracellular Matrix/metabolism , Gene Expression Regulation , Humans , Intervertebral Disc Degeneration/pathology , Middle Aged , Phenotype
6.
Eur Cell Mater ; 28: 98-110; discussion 110-1, 2014 Sep 12.
Article in English | MEDLINE | ID: mdl-25214017

ABSTRACT

Excessive mechanical loading or acute trauma to intervertebral discs (IVDs) is thought to contribute to degeneration and pain. However, the exact mechanisms by which mechanical injury initiates and promotes degeneration remain unclear. This study investigates biochemical changes and extracellular matrix disruption in whole-organ human IVD cultures following acute mechanical injury. Isolated healthy human IVDs were rapidly compressed by 5% (non-injured) or 30% (injured) of disc height. 30% strain consistently cracked cartilage endplates, confirming disc trauma. Three days post-loading, conditioned media were assessed for proteoglycan content and released cytokines. Tissue extracts were assessed for proteoglycan content and for aggrecan integrity. Conditioned media were applied to PC12 cells to evaluate if factors inducing neurite growth were released. Compared to controls, IVD injury caused significant cell death. Injury also caused significantly reduced tissue proteoglycan content with a reciprocal increase of proteoglycan content in culture media. Increased aggrecan fragmentation was observed in injured tissue due to increased matrix metalloproteinase and aggrecanase activity. Injured-IVD conditioned media contained significantly elevated interleukin (IL)-5, IL-6, IL-7, IL-8, MCP-2, GROα, and MIG, and ELISA analysis showed significantly increased nerve growth factor levels compared to non-injured media. Injured-disc media caused significant neurite sprouting in PC12 cells compared to non-injured media. Acute mechanical injury of human IVDs ex vivo initiates release of factors and enzyme activity associated with degeneration and back pain. This work provides direct evidence linking acute trauma, inflammatory factors, neo-innervation and potential degeneration and discogenic pain in vivo.


Subject(s)
Extracellular Matrix Proteins/metabolism , Intervertebral Disc Degeneration/etiology , Intervertebral Disc/metabolism , Stress, Mechanical , Adult , Cell Death , Culture Media, Conditioned/pharmacology , Cytokines/metabolism , Fractures, Cartilage/complications , Fractures, Cartilage/metabolism , Humans , Intervertebral Disc/injuries , Intervertebral Disc Degeneration/metabolism , Middle Aged , Neurites/drug effects , Pain/etiology , Pain/metabolism
7.
J Hum Hypertens ; 25(12): 732-8, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21228822

ABSTRACT

Failing to reach blood pressure (BP) goals is one of the main problems in hypertension management. Especially in high-risk patients, intensive monitoring including frequently office visits or new techniques to monitor home BP is required. A total of 60 patients with uncontrolled hypertension were included and randomized into a group with telemetric BP monitoring (TBPM) (n=30) and a control group receiving standard care (n=30). During the 3-month study period, patients received in addition to their antihypertensive pre-treatment up to 2 × 300 mg irbesartan to achieve the required target BP. All patients were instructed to measure their BP once daily in the morning. In the TBPM group automatic alerts were generated by the central database server using pre-defined algorithms and patients were subsequently contacted by the physician. At baseline mean 24-h ambulant BP monitoring (ABPM) was 143.3±11.1/82.6±9.9 mm Hg in the TBPM group and 141.4±12.6/82.1±6.5 mm Hg in the standard care group. During treatment mean systolic BP showed a more intensive decrease in the TBPM vs control group (-17.0±11.1 mm Hg vs -9.8±13.7 mm Hg; P=0.032). Patients in the TBPM group had a more pronounced night dipping and a higher reduction of mean pulse pressure than controls (-8.1±5.9 mm Hg vs -2.8±7.4 mm Hg, P=0.004). After 3 months, TBPM-treated patients were given a higher mean daily dose of irbesartan (375±187 mg vs 222±147 mg in controls; P=<0.001). We demonstrated that with TBPM a more effective and faster titration of the antihypertensive agent is possible. The alarm criteria chosen were useful to improve BP control.


Subject(s)
Antihypertensive Agents/therapeutic use , Biphenyl Compounds/therapeutic use , Blood Pressure Monitoring, Ambulatory/methods , Blood Pressure/physiology , Hypertension/drug therapy , Monitoring, Ambulatory/methods , Telemedicine/methods , Tetrazoles/therapeutic use , Adult , Aged , Algorithms , Antihypertensive Agents/pharmacology , Biphenyl Compounds/pharmacology , Blood Pressure/drug effects , Circadian Rhythm/physiology , Female , Humans , Hypertension/physiopathology , Irbesartan , Male , Middle Aged , Office Visits , Physician-Patient Relations , Tetrazoles/pharmacology , Time Factors , Treatment Outcome
8.
Phys Rev Lett ; 99(8): 085502, 2007 Aug 24.
Article in English | MEDLINE | ID: mdl-17930954

ABSTRACT

This study revealed the nature of native defects and their roles in ZnO through positron annihilation and optical transmission measurements. It showed oxygen vacancies are the origin for the shift in the optical absorption band that causes the red or orange coloration. It also revealed experimental evidence that the donor nature of oxygen vacancy is approximately 0.7 eV. In addition, this work showed the Zn interstitial was not the donor in the as-grown ZnO and supported recent calculations that predicted hydrogen in an oxygen vacancy forms multicenter bonds and acts as a shallow donor.


Subject(s)
Electrons , Zinc Oxide , Oxygen , Zinc Oxide/chemistry
9.
Dtsch Med Wochenschr ; 132(9): 423-6, 2007 Mar 02.
Article in German | MEDLINE | ID: mdl-17315118

ABSTRACT

BACKGROUND AND OBJECTIVE: Non-adherence to fluid intake restrictions is one of the leading problems in hemodialysis patients. The consequences of chronic volume overload and massive hypotensive episodes resulting from enhanced ultrafiltration lead to an increased mortality and incidence of vascular events. Telemetric body weight monitoring (TBWM) suggests itself as a successful way to reduce daily fluid intake PATIENTS AND METHODS: This monocentric, prospective, randomized open study includes 120 patients with end-stage renal failure undergoing chronic hemodialysis (for at least two months) three times a week. The mean interdialytic weight gain (IWG) was more than 1.5 kg/2 days over the four weeks immediately before start of the study. The effect of daily body weight telemonitoring on IWG, blood pressure, haemoglobin variability, hospital stay, vascular events and mortality were observed for three months. All monitored patients (group 1, n = 60) received a weekly report of their weight changes, the number of alarms (automatically sent by email to the study center when daily IWG was greater than 0.75 kg/d) and of the interventions by phone (conducted by the responsible nephrologist when IWG was > 2 kg/day). Hemodynamics (each hemodialysis procedure) and weekly laboratory data were recorded for all patients. RESULTS: Preliminary data of 44 patients showed a significant reduction of daily IWG (weekly average, p = 0.0187) and a smaller number of alarm reports after the whole study period in group 1. Blood pressure monitoring during hemodialysis showed less hyper- and hypotensive episodes in patients with an IWG of less than 1.5 kg/2 days. In the control group there have so far been no changes of the analysed parameters. CONCLUSIONS: TBWM seems to be an effective method for optimizing adherence to fluid intake restrictions in patients on hemodialysis. Hemoglobin variability, mortality rates and the number of vascular events will still have to be analysed in detail for all patients once the entire study period has been completed.


Subject(s)
Body Weight , Kidney Failure, Chronic/therapy , Renal Dialysis , Telemetry/methods , Blood Pressure , Hemoglobins/analysis , Humans , Kidney Failure, Chronic/mortality , Kidney Failure, Chronic/physiopathology , Monitoring, Physiologic/methods , Prospective Studies , Survival Analysis , Terminal Care , Water-Electrolyte Balance , Weight Gain
10.
J Bacteriol ; 183(24): 7381-6, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11717297

ABSTRACT

The cold shock response in both Escherichia coli and Bacillus subtilis is induced by an abrupt downshift in growth temperature and leads to a dramatic increase in the production of a homologous class of small, often highly acidic cold shock proteins. This protein family is the prototype of the cold shock domain (CSD) that is conserved from bacteria to humans. For B. subtilis it has been shown that at least one of the three resident cold shock proteins (CspB to D) is essential under optimal growth conditions as well as during cold shock. Analysis of the B. subtilis cspB cspC double deletion mutant revealed that removal of these csp genes results in pleiotropic alteration of protein synthesis, cell lysis during the entry of stationary growth phase, and the inability to differentiate into endospores. We show here that heterologous expression of the translation initiation factor IF1 from E. coli in a B. subtilis cspB cspC double deletion strain is able to cure both the growth and the sporulation defects observed for this mutant, suggesting that IF1 and cold shock proteins have at least in part overlapping cellular function(s). Two of the possible explanation models are discussed.


Subject(s)
Bacillus subtilis/genetics , Bacterial Proteins , Carrier Proteins/genetics , Escherichia coli Proteins , Escherichia coli/genetics , Eukaryotic Initiation Factor-1/genetics , Heat-Shock Proteins/genetics , Cold Temperature , Eukaryotic Initiation Factor-1/chemistry , Gene Expression Regulation, Bacterial , Genetic Complementation Test , Models, Molecular , Molecular Chaperones , RNA, Bacterial/metabolism , RNA-Binding Proteins , Signal Transduction
11.
J Bacteriol ; 183(21): 6435-43, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11591689

ABSTRACT

Using immunofluorescence microscopy and a fusion of a cold shock protein (CSP), CspB, to green fluorescent protein (GFP), we showed that in growing cells Bacillus subtilis CSPs specifically localize to cytosolic regions surrounding the nucleoid. The subcellular localization of CSPs is influenced by the structure of the nucleoid. Decondensed chromosomes in smc mutant cells reduced the sizes of the regions in which CSPs localized, while cold shock-induced chromosome compaction was accompanied by an expansion of the space in which CSPs were present. As a control, histone-like protein HBsu localized to the nucleoids, while beta-galactosidase and GFP were detectable throughout the cell. After inhibition of translation, CspB-GFP was still present around the nucleoids in a manner similar to that in cold-shocked cells. However, in stationary-phase cells and after inhibition of transcription, CspB was distributed throughout the cell, indicating that specific localization of CspB depends on active transcription and is not due to simple exclusion from the nucleoid. Furthermore, we observed that nucleoids are more condensed and frequently abnormal in cspB cspC and cspB cspD double-mutant cells. This suggests that the function of CSPs affects chromosome structure, probably through coupling of transcription to translation, which is thought to decondense nucleoids. In addition, we found that cspB cspD and cspB cspC double mutants are defective in sporulation, with a block at or before stage 0. Interestingly, CspB and CspC are depleted from the forespore compartment but not from the mother cell. In toto, our findings suggest that CSPs localize to zones of newly synthesized RNA, coupling transcription with initiation of translation.


Subject(s)
Bacillus subtilis/chemistry , Bacillus subtilis/genetics , Bacterial Proteins/analysis , Bacillus subtilis/physiology , Bacterial Proteins/genetics , Bacterial Proteins/physiology , Cell Division , Cell Nucleus/ultrastructure , Chromosome Segregation , Cytosol/chemistry , Gene Expression Regulation, Bacterial , Green Fluorescent Proteins , Luminescent Proteins/genetics , Microscopy, Fluorescence , Mutation , Protein Biosynthesis , Recombinant Fusion Proteins/analysis , Recombinant Fusion Proteins/physiology , Spores, Bacterial/genetics , Transcription, Genetic
12.
Neurochem Res ; 26(2): 161-6, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11478743

ABSTRACT

We have studied the developmental changes of glucose, mannose, fructose and galactose metabolism in rat cerebral cortex. As the animals aged, glucose, mannose and fructose oxidation to CO2 increased, whereas galactose oxidation decreased. Lipid synthesis from glucose and fructose also increased with age, that from mannose decreased and galactose did not change. Cytochalasin B, a potent non-competitive inhibitor of sodium-independent glucose transport, significantly impaired glucose, mannose and galactose metabolism, but had no effect on fructose metabolism. Both galactose or fructose did not change, whereas mannose declined the glucose metabolism. Glucose decreased fructose, galactose and mannose metabolism. Our results show that besides glucose, the metabolism of mannose, galactose and fructose present developmental changes from fetal to adult age, and reinforce the literature data indicating that mannose and galactose are transported by glucose carriers, while fructose is not.


Subject(s)
Aging/metabolism , Cerebral Cortex/enzymology , Hexoses/metabolism , Animals , Carbon Dioxide/metabolism , Cerebral Cortex/drug effects , Cerebral Cortex/growth & development , Cytochalasin B/pharmacology , Dose-Response Relationship, Drug , Lipids/biosynthesis , Oxidation-Reduction/drug effects , Rats , Rats, Wistar
13.
EMBO Rep ; 2(8): 685-9, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11463749

ABSTRACT

The large subunit of ribosomes in Bacillus subtilis was tagged by generation of a fusion of ribosomal protein L1 to blue fluorescent protein (BFP). The fusion was fully active and localized around the nucleoids, predominantly close to the cell poles, in growing cells. However, in stationary phase cells, and in growing cells treated with rifampicin, L1-BFP was distributed throughout the cells, in contrast to cells treated with chloramphenicol, in which ribosomes still localized around nucleoids. These data show that specific localization of ribosomes is not due to nucleoid exclusion, but is a dynamic process due to active synthesis of RNA. Dual labelling of ribosomes and cold shock proteins (CSPs) tagged with green fluorescent protein (GFP) revealed colocalization of both protein classes. CSPs are implicated in coupling of transcription with translation and may bridge the spatial separation of ribosomes and nucleoid-associated RNA polymerase.


Subject(s)
Bacillus subtilis/metabolism , Ribosomal Proteins/metabolism , Ribosomes/metabolism , Transcription, Genetic , Bacillus subtilis/genetics , Bacillus subtilis/ultrastructure , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Genes, Reporter , Green Fluorescent Proteins , Heat-Shock Proteins/genetics , Heat-Shock Proteins/metabolism , Indicators and Reagents/metabolism , Luminescent Proteins/metabolism , Microscopy, Fluorescence , Models, Biological , Protein Subunits , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Replication Origin , Ribosomal Proteins/genetics
14.
Mol Microbiol ; 39(5): 1321-9, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11251847

ABSTRACT

In our attempt to understand the cold shock response of Bacillus subtilis, we report on the role of the B. subtilis fatty acid desaturase (FA-D) Des during membrane adaptation to low temperatures and demonstrate its importance during cold shock. A des null mutant was constructed and analysed in comparison with its parental strain. Growth studies and large-scale comparative fatty acid (FA) analysis revealed a severe cold-sensitive phenotype of the des deletion mutant during the absence of isoleucine and showed that four unsaturated fatty acid (UFA) species differing in length, branching pattern and position of the double bond are synthesized in B. subtilis JH642 but not in the des null mutant. Apart from the lack of UFA synthesis, the FA-D deletion strain showed a dramatically altered saturated fatty acid (SFA) profile at the onset of the stationary growth phase in the presence of exogenous isoleucine sources. Expression of des integrated in trans at the amyE locus of the des deletion strain not only cured the cold-sensitive phenotype observed for the des mutant but allowed much better growth than in strain JH642 after a shift from 37 degrees C to 15 degrees C. These results show that, during cold shock adaptation, des expression can completely replace the isoleucine-dependent, long-term, FA branching adaptation mechanism. We conclude that the crucial aspect in cold adaptation of the cytoplasmic membrane is not its specific molecular composition but rather its physical status in terms of its fluidity.


Subject(s)
Adaptation, Physiological , Bacillus subtilis/physiology , Cell Membrane/physiology , Cold Temperature , Fatty Acid Desaturases/physiology , Bacillus subtilis/chemistry , Bacillus subtilis/genetics , Cell Membrane/chemistry , Culture Media , Fatty Acids/analysis , Gene Deletion , Heat-Shock Response/physiology
15.
J Hematother Stem Cell Res ; 9(6): 861-5, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11177598

ABSTRACT

Acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) is observed in 5 to 10% of patients treated with high-dose chemotherapies followed by autologous stem cell and bone marrow transplantation. Both diseases are frequently associated with monosomy 7 (-7), trisomy 8 (+8), loss of the long arm of chromosome 5 (-5q), and deletions including the TP53-gene region according to del(17)(p13). In this study, we examined whether these chromosomal aberrations are already detectable in blood stem cells from patients who have all been treated with standard chemotherapies prior to peripheral blood stem cell transplantation (PBSCT). Therefore, we screened peripheral blood derived stem cells obtained at the time of stem cell harvest for the presence of -7, +8, -5q, and del(17)(p13) by fluorescence in situ hybridization (FISH). Our series included 40 patients: 4 patients with Hodgkin's disease, 6 patients with non-Hodgkin-lymphoma (NHL), 1 patient with ALL, 4 patients with plasmocytoma, and 25 patients with solid tumors. Peripheral blood mononuclear cells (PBMC) from eight healthy blood donors served as controls. Assuming a hybridization efficiency of >98%, the cut-off level of non diploid cells was determined for each DNA-probe. None of the stem cell preparations exhibited chromosomal damage. Our findings indicate that chromosomal damage is a rare event in stem cell autografts.


Subject(s)
Chromosome Aberrations/diagnosis , Hematopoietic Stem Cell Transplantation/standards , Hematopoietic Stem Cells/metabolism , Leukemia, Myeloid/genetics , Myelodysplastic Syndromes/genetics , Acute Disease , Adult , Case-Control Studies , Chromosome Aberrations/genetics , Chromosome Disorders , Cohort Studies , Female , Hematopoietic Stem Cell Transplantation/methods , Humans , In Situ Hybridization, Fluorescence , Leukapheresis , Leukemia, Myeloid/blood , Leukemia, Myeloid/therapy , Male , Middle Aged , Myelodysplastic Syndromes/blood , Myelodysplastic Syndromes/therapy , Neoplasms/blood , Neoplasms/genetics , Neoplasms/therapy , Transplantation, Autologous/methods , Transplantation, Autologous/standards
16.
Crit Rev Eukaryot Gene Expr ; 10(3-4): 281-302, 2000.
Article in English | MEDLINE | ID: mdl-11272470

ABSTRACT

Bone metastases occur in approximately 80% of patients with advanced cancer. They are characterized by cancer cell growth and bone destruction that cause pain, fractures, anemia, and hypercalcemia. At diagnosis, bone metastases are usually incurable owing to their advanced development. However, the early stages in their formation are asymptomatic and begin as single micrometastatic cells from the blood stream. These cells can be detected by molecular analysis of bone marrow in approximately 30% of patients at the time of cancer diagnosis, but not all single micrometastatic cells develop into clinically significant bone metastases. A synergistic relationship exists between the micometastasis and the bone environment creating favorable conditions for the development and growth of disseminated tumor cells. Such bone metastases induce osteolysis or new bone formation, releasing growth factors and cytokines, which in turn amplify this pathological mechanism. The underling hypothesis, first proposed by Paget in 1889, is that the growth of disseminated tumor cells in bone is dependent on the fertility of the soil or bone itself. This article explores the most current opinions in this area of study and presents a comprehensive summary of the major factors involved.


Subject(s)
Bone Neoplasms/secondary , Bone Neoplasms/pathology , Cell Adhesion , Cell Division , Cell Movement , Chemotaxis , Humans , Neoplasm Seeding , Neovascularization, Pathologic , Osteoblasts/cytology , Osteolysis
17.
J Bacteriol ; 181(17): 5341-9, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10464205

ABSTRACT

Bacillus subtilis has developed sophisticated mechanisms to withstand fluctuations in temperature. Membrane fatty acids are the major determinants for a sufficiently fluid membrane state to ensure the membrane's function at all temperatures. The fatty acid profile of B. subtilis is characterized by a high content of branched fatty acids irrespective of the growth medium. Here, we report on the importance of isoleucine for B. subtilis to survive cold shock from 37 to 15 degrees C. Cold shock experiments with strain JH642 revealed a cold-protective function for all intermediates of anteiso-branched fatty acid biosynthesis. Metabolites related to iso-branched or straight-chain fatty acid biosynthesis were not protective. Fatty acid profiles of different B. subtilis wild-type strains proved the altered branching pattern by an increase in the anteiso-branched fatty acid content and a concomitant decrease of iso-branched species during cold shock. There were no significant changes in the fatty acid saturation or acyl chain length. The cold-sensitive phenotype of isoleucine-deficient strains in the absence of isoleucine correlated with their inability to synthesize more anteiso-branched fatty acids, as shown by the fatty acid profile. The switch to a fatty acid profile dominated by anteiso-C(15:0) and C(17:0) at low temperatures and the cold-sensitive phenotype of isoleucine-deficient strains in the absence of isoleucine focused our attention on the critical role of anteiso-branched fatty acids in the growth of B. subtilis in the cold.


Subject(s)
Adaptation, Physiological , Bacillus subtilis/metabolism , Bacillus subtilis/physiology , Fatty Acids/metabolism , Isoleucine/metabolism , Bacillus subtilis/growth & development , Biological Transport , Cell Membrane/physiology , Cold Temperature , Kinetics , Phenotype , Threonine/metabolism
18.
Mol Microbiol ; 25(4): 741-56, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9379903

ABSTRACT

Like other bacteria, Bacillus subtilis possesses a family of homologous small acidic proteins (CspB, CspC and CspD, identity >70%) that are strongly induced in response to cold shock. We show that deletion of cspC or cspD genes did not result in a detectable phenotype; in contrast, csp double mutants exhibited severe reduction in cellular growth at 15 degrees C as well as at 37 degrees C, including impairment of survival during the stationary phase. Two-dimensional gel analysis showed that protein synthesis was deregulated in csp double mutants and that the loss of one or two CSPs led to an increase in the synthesis of the remaining CSP(s) at 37 degrees C and after cold shock, suggesting that CSPs down-regulate production of members from this protein family. A cspB/C/D triple mutant (64BCDbt) could only be generated in the presence of cspB in trans on a plasmid that was not lost, in spite of lack of antibiotic pressure, indicating that a minimum of one csp gene is essential for viability of B. subtilis. After cold shock, synthesis of CspB in 64BCDbt was drastically lower than in wild-type cells accompanied by cessation in growth and strong reduction in general protein synthesis. As CspB, CspC and CspD are shown to bind to RNA in a co-operative and interactive manner, CSPs are suggested to function as RNA chaperones facilitating the initiation of translation under optimal and low temperatures.


Subject(s)
Bacillus subtilis/physiology , Cold Temperature , Heat-Shock Proteins/physiology , Bacillus subtilis/genetics , Bacterial Proteins/biosynthesis , Bacterial Proteins/physiology , Base Sequence , Cell Division/physiology , Cloning, Molecular , Gene Deletion , Gene Expression Regulation, Bacterial/genetics , Genes, Bacterial/genetics , Heat-Shock Proteins/genetics , Molecular Sequence Data , Sequence Alignment , Sequence Analysis, DNA
19.
Urologe A ; 36(1): 68-76, 1997 Jan.
Article in German | MEDLINE | ID: mdl-9123685

ABSTRACT

In 180 children (87 children belonging to a control group, 68 with fever of non-renal origin, and 25 with pyelonephritis) albumin and immunoglobulin G (markers for glomerular dysfunction), alpha-1-microglobulin and beta-NAG (markers for proximal tubular dysfunction) and apolipoprotein A1 (marker of "postrenal' dysfunction) were measured in second-voided morning urine. In children with fever of non-renal origin, glomerular dysfunction was encountered in 8.8%, tubular dysfunction in 17.6% and mixed glomerular-tubular dysfunction in 14.7% of cases. Among children with pyelonephritis, 28% revealed glomerular dysfunction and 44% mixed glomerular-tubular dysfunction. No case of solitary proximal tubular dysfunction was observed in children with pyelonephritis. There were highly significant differences in presence and expression of glomerular dysfunction between children with fever of non-renal origin and children with pyelonephritis (P < 0.0001), whereas with regard to proximal tubular dysfunction, the differences were only moderately significant (beta-NAG: P < 0.01) or of low significance (alpha-1-microglobulin: P < 0.05). This may indicate that morphologic changes occur during interstitial pyelonephritis due to inflammation of glomeruli, resulting in glomerular dysfunction, while proximal tubular dysfunction may additionally be due to fever-associated function processes.


Subject(s)
Enzymes/urine , Fever of Unknown Origin/etiology , Proteinuria/diagnosis , Pyelonephritis/diagnosis , Acetylglucosaminidase/urine , Adolescent , Albuminuria/diagnosis , Albuminuria/urine , Apolipoprotein A-I/urine , Child , Child, Preschool , Diagnosis, Differential , Female , Fever of Unknown Origin/urine , Humans , Immunoglobulin G/urine , Infant , Kidney Function Tests , Kidney Glomerulus/physiopathology , Kidney Tubules/physiopathology , Male , Prospective Studies , Proteinuria/urine , Pyelonephritis/urine
20.
Z Arztl Fortbild (Jena) ; 90(3): 227-31, 1996 May.
Article in German | MEDLINE | ID: mdl-8779236

ABSTRACT

40% of the male patients suffering from gouty arthritis treated in our rheumatological unit during a three years period showed a chronic polyarticular course. In polyarticular gout, acute gouty attacks affect above all the joints of the upper limbs. Gouty arthritis in these patients shows an ascending pattern. Therefore, in case of unawareness of the clinical symptoms, differential diagnosis can be difficult especially when laboratory findings including uric acid levels are in normal ranges and classical radiological findings are missing. It is important to notice that the atypical joint attack in gouty arthritis is typical for polyarticular gout. In case of unclear arthritis the polarized light microscopy of the synovial fluid should always be demanded.


Subject(s)
Arthritis, Gouty/diagnosis , Adult , Aged , Arthritis, Gouty/blood , Arthritis, Gouty/therapy , Diagnosis, Differential , Follow-Up Studies , Humans , Joints/pathology , Male , Microscopy, Polarization , Middle Aged , Synovial Fluid/chemistry , Uric Acid/blood
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