Impact of 5 fluorouracil chemotherapy on gut inflammation, functional parameters, and gut microbiota.

Emerging evidence suggests that gut microbiota may influence the response to chemotherapy. We sought to characterize the effects of 5 fluorouracil (5FU) chemotherapy on colon inflammation and functional measures in colorectal cancer (CRC) and to further determine whether gut microbiota can influence this response. 50 C57BL/6 were randomized into four groups; Control + Vehicle (n = 10), Control + 5FU (n = 10), AOM/DSS + Vehicle (n = 15), and AOM/DSS + 5FU (n = 15). CRC was induced chemically by a single 10 mg/kg injection of azoxymethane (AOM) followed by two cycles (2% and 1%) of dextran sodium sulfate (DSS). Mice were then treated with 3 cycles of vehicle or 5FU (cycle 1: 40 mg/kg, cycle 2 + 3: 20 mg/kg). Functional tests (grip strength and run-to-fatigue) were performed prior to 5FU treatment (baseline) and at the completion of the second cycle of 5FU. Following the third 5FU cycle, mice were euthanized and the colon was evaluated for expression of inflammatory genes using RT-qPCR and stool samples were profiled using 16S rRNA sequencing. A second experiment used fecal microbiota transplantation from 5FU treated mice to control mice (n = 10-15/group) to determine whether 5FU associated changes in the microbiota could influence functional measures and colon inflammation. 5FU reduced grip strength (p < 0.05) and caused a trending decrease in run-to-fatigue performance in cancer mice (p = 0.06). Select intestinal inflammatory genes were significantly elevated with 5FU treatment and this was further exacerbated with cancer (p < 0.05). Microbiota analysis revealed increased dissimilarity and alterations in bacterial taxonomy in 5FU and AOM/DSS-treated mice (p < 0.05). Fecal transplant from 5FU treated mice reduced functional performance (p < 0.05) and altered select colon inflammatory markers (p < 0.05). This study provides evidence of an effect of 5FU on inflammatory responses and functional measures in a mouse model of CRC and suggests that gut microbes may play a role in some, but not all, 5FU related perturbations.

Effects of 5-fluorouracil chemotherapy on fatigue: role of MCP-1.

Chemotherapy has been known to cause severe side effects, including fatigue. While the mechanisms for chemotherapy induced fatigue (CIF) are likely to be multi-factorial in origin, it is thought that inflammation and anemia may play a role. The purpose of this study was to examine the effect of chemotherapy on fatigue in mice, and further, to begin to determine if inflammation and anemia may contribute to this response. For experiment 1, C57BL/6 mice were assigned to: vehicle (PBS), low (20 mg/kg), medium (40 mg/kg), or high (60 mg/kg) doses of 5-fluorouracil (5-FU). Voluntary physical activity (PA) was measured throughout the treatment period (day 1-5) as well as during the recovery period (day 6-14). In experiment 2, we examined the effects of 5-FU (60 mg/kg) on the inflammatory mediator MCP-1 and on markers of anemia (RBC, Hct and Hb). Finally, using MCP-1(-/-) mice we examined the role of MCP-1 on CIF (experiment 3). 5-FU reduced voluntary PA in a dose response manner (p<0.05). Plasma MCP-1 was increased following 5-FU treatment on both days 5 (p=0.10) and 14 (p<0.05). In addition, RBCs, Hct and Hb were reduced with 5-FU on days 5 and 14 (p<0.05). Both C57BL/6 and MCP-1(-/-) mice saw similar decrements in PA through the duration of the treatment period (days 1-5), however the MCP-1(-/-) mice recovered much earlier than wildtype mice. This study provides evidence of the dose response effect of a standard chemotherapy agent on fatigue and demonstrates a potential role of MCP-1 and presumably inflammation, and anemia.

Benefits of exercise training on breast cancer progression and inflammation in C3(1)SV40Tag mice.

Many observational epidemiologic studies suggest an association between exercise and breast cancer risk. However, the lack of controlled experimental studies that examine this relationship and the mechanisms involved weaken the basis for inferring a causal relationship. Inflammation plays a role in breast cancer progression and exercise has been reported to reduce inflammation; however, the anti-inflammatory effects of exercise in breast cancer have yet to be established. We examined the relationship between exercise training and systemic inflammation in relation to breast cancer progression in C3(1)SV40Tag mice. Female C3(1)SV40Tag mice were assigned to either exercise (Ex) or sedentary (Sed) treatment (n=12-14/group). Beginning at 4 wks of age mice (Ex) were run on a treadmill for 60 min/d (20 m/min and 5% grade), 6 d/wk for a period of 20 wks. Mice were examined weekly for palpable tumors, and tumor number and volume were recorded. At 24 wks of age mice were sacrificed and a more direct measure of tumor number and volume, and spleen weight was recorded. Plasma was analyzed for MCP-1 and IL-6 concentration using ELISA. Ex reduced palpable tumor number at sacrifice (24 wks) by approximately 70% (P<0.05). Tumor volume was also reduced in Ex at 21-23 wks (P<0.05). This reduction in tumor progression by Ex was associated with a reduction in plasma concentration of MCP-1 and IL-6, and spleen weight (P<0.05). These data provide strong support for a beneficial effect of exercise training on tumor progression in the C3(1)SV40Tag mouse model of breast cancer that may be partly mediated by its anti-inflammatory potential.

Neuroradiologic correlation with aphasias. Cortico-subcortical map of language. / Correlación neurorradiológica de las afasias. Mapa cortico-subcortical del lenguaje.

Aphasia is an acquired language disorder due to a cerebral lesion; it is characterized by errors in production, denomination, or comprehension of language. Although most aphasias are mixed, from a practical point of view they are classified into different types according to their main clinical features: Broca's aphasia, Wernicke's aphasia, conduction aphasia, transcortical aphasia, and alexia with or without agraphia. We present the clinical findings for the main subtypes of aphasia, illustrating them with imaging cases, and we provide an up-to-date review of the language network with images from functional magnetic resonance imaging and tractography.

Dynamic regulation of copper uptake and detoxification genes in Saccharomyces cerevisiae.

The essential yet toxic nature of copper demands tight regulation of the copper homeostatic machinery to ensure that sufficient copper is present in the cell to drive essential biochemical processes yet prevent the accumulation to toxic levels. In Saccharomyces cerevisiae, the nutritional copper sensor Mac1p regulates the copper-dependent expression of the high affinity Cu(I) uptake genes CTR1, CTR3, and FRE1, while the toxic copper sensor Ace1p regulates the transcriptional activation of the detoxification genes CUP1, CRS5, and SOD1 in response to copper. In this study, we characterized the tandem regulation of the copper uptake and detoxification pathways in response to the chronic presence of elevated concentrations of copper ions in the growth medium. Upon addition of CuSO4, mRNA levels of CTR3 were rapidly reduced to eightfold the original basal level whereas the Ace1p-mediated transcriptional activation of CUP1 was rapid and potent but transient. CUP1 expression driven by an Ace1p DNA binding domain-herpes simplex virus VP16 transactivation domain fusion was also transient, demonstrating that this mode of regulation occurs via modulation of the Ace1p copper-activated DNA binding domain. In vivo dimethyl sulfate footprinting analysis of the CUP1 promoter demonstrated transient occupation of the metal response elements by Ace1p which paralleled CUP1 mRNA expression. Analysis of a Mac1p mutant, refractile for copper-dependent repression of the Cu(I) transport genes, showed an aberrant pattern of CUP1 expression and copper sensitivity. These studies (i) demonstrate that the nutritional and toxic copper metalloregulatory transcription factors Mac1p and Ace1p must sense and respond to copper ions in a dynamic fashion to appropriately regulate copper ion homeostasis and (ii) establish the requirement for a wild-type Mac1p for survival in the presence of toxic copper levels.

Essential Subcortical Tracts in Language and Reading. 3D-Tractography for Clinical Practice and Anatomic Correlation with Intraoperative Subcortical Electrostimulation.

Diffusion tensor imaging (DTI) and tractography provide the neurosurgeon with a valid 3D view of the white matter tracts of the brain for the presurgical planning of the treatment of lesions close to eloquent areas, this being one of the principal clinical applications of this technique. In this article, we describe through practical cases the anatomic relationships of white matter tracts that are essential for language and reading, based on DTI studies and the excellent anatomic correlation with the intraoperative subcortical map.

Copper-binding motifs in catalysis, transport, detoxification and signaling.

Copper is required for many biological processes but is toxic at high cellular concentrations, so levels in the cell must be strictly controlled. Copper-binding motifs have been identified and characterized in many proteins. The way in which copper is coordinated by these motifs is important for the transport and distribution of intracellular copper and for the effective functioning of copper-dependent enzymes.

Isolation and culture of human endometrial glands.

A simple method for isolation of glands from human endometrium has been developed. The procedure involves collagenase digestion of the endometrial tissue and filtration through sieves of various pore sizes. Isolated glands retained on the sieves were washed and collected in culture dishes. Tubular organization of the isolated glands was ascertained by examination of the preparations under inverted microscope and light microscopy of stained sections. The appearance of the glands was found to reflect different functional states of the endometrium and, possibly, to reveal abnormalities. Growth of monolayers of epithelial cells derived from the glands was observed within 24 h of culturing. Electron microscopy of the cells in 7-day monolayer preparations from both proliferative and secretory endometrium revealed the characteristic features of human endometrial epithelial cells, viz. presence of microvilli and desmosome-like junctions. Nuclear bodies were observed in cells derived from both types of endometrium.

Beneficial effects of increasing monounsaturated fat intake in adolescents with type 1 diabetes.

This study aimed to increase the monounsaturated fat content in the diet of outpatient adolescents with type 1 diabetes and to examine the metabolic effects after 12 weeks. Twenty-three adolescents were randomly allocated to either a high monounsaturated fat diet or a control diet. Their mean age was 16.9 (S.D. 2.1) years and median HbA(1c) was 9.1% [IQR 7.9-10.4%]. Dietary targets were not reached judged by their 4-day food diaries. However, the whole study group had a significant increase in monounsaturated fat as indexed by red cell phospholipid fatty acids (RCFAs), with an increase of n-9 RCFAs from 14.9% [IQR: 14.5-21.7%] to 21.7% [IQR: 18.8-25.6%] (P=0.002). Changes in n-9 RCFAs were inversely related to changes in HbA(1c) (R(2)=0.26, P=0.02), such that a 10% increase in n-9 RCFAs corresponded to a 0.64% improvement (decrease) in HbA(1c). Changes in n-9 RCFAs were also inversely related to changes in plasma total cholesterol (R(2)=0.38, P=0.002) and plasma LDL cholesterol (R(2)=0. 21, P=0.03). These changes were not associated with changes in insulin dose, body weight or physical activity. Overall, the results demonstrate that a modest increase in the monounsaturated fat content of an adolescent diet has the potential to improve glycaemic control and lipid profile.

Vascular responses by transcutaneous oximetry in adolescents with and without diabetes.

The vascular response of the skin was evaluated by transcutaneous oximetry (TcPO2) in the forearm in 119 adolescents with type I diabetes aged 10.4-19.8 (median 15.3) years, with a duration of diabetes 0.7 to 18.3 (median 7.8) years, and 49 nondiabetic adolescents aged 11.3-18.8 (median 15.5) years. Two different vascular stimuli were used: heating of the probe to 43 degrees C and 5 min of ischemia. Baseline TcPO2 after 13 min of equilibration at a probe temperature of 43 degrees C, postischemic maximum TcPO2, and the postischemic TcPO2 increase were significantly lower in the diabetic group compared to the control group (p = 0.0001, p < 0.0001, and p = 0.0001, respectively). In both the diabetic and the control groups, gender differences were found for baseline TcPO2 (p = 0.0001 and p = 0.0009, respectively) and postischemic maximum TcPO2 (p = 0.0001 and p = 0.005, respectively), the girls having consistently higher values. After controlling for gender by multiple linear regression analysis, duration of diabetes showed a significant effect on postischemic maximum TcPO2 (R2 = 22%, p = 0.02). The postischemic TcPO2 increase was not affected by gender. Lower values for the postischemic TcPO2 increase were related to higher GHb values (R2 = 4%, p = 0.03). Abnormal values for oximetry were associated only with some autonomic nerve function abnormalities. Differences in the vascular response to heat and ischemia as measured by transcutaneous oximetry can be demonstrated between adolescents with type I diabetes and nondiabetic controls, as well as between girls and boys. Lower values in diabetic subjects are weakly associated with diabetes duration and metabolic control, independent of gender.

Supplementation with flaxseed oil versus sunflowerseed oil in healthy young men consuming a low fat diet: effects on platelet composition and function.

OBJECTIVE: To compare the effects of supplementing a low fat diet with an alpha-linolenic acid-rich (C18:3 n-3) oil with a linoleic acid-rich (C18:2 n-6) oil on platelet composition and function. DESIGN: Prospective study with random allocation to one of the two oils. SETTING: Free-living study. SUBJECTS: Eleven healthy young males recruited from within the University. INTERVENTIONS: Subjects were allocated to consume 40 g of either flaxseed oil (n = 5) or sunflowerseed oil (n = 6) daily for 23 days. Fasting blood samples were collected at commencement and completion of supplementation for analysis of platelet fatty acids and platelet aggregation. RESULTS: The platelet eicosapentaenoic acid (EPA) more than doubled in the group taking flaxseed oil (P < 0.05) but was unchanged in the sunflowerseed group. As a result the platelet EPA:arachidonic acid ratio, considered a marker for thromboxane production and platelet aggregation potential, increased in the flaxseed group (P < 0.05). The aggregation response induced by 0.75 and 2 micrograms of collagen was decreased in those taking flaxseed oil (P < 0.05). CONCLUSION: This study provides further evidence that consumption of alpha-linolenic acid-rich oils may offer protective effects against cardiovascular disease over linoleic acid-rich oils via their ability to decrease the tendency of platelets to aggregate.

Biodegradation of phenol in a continuous process: comparative study of stirred tank and fluidized-bed bioreactors.

The paper presents the main results obtained from the study of the biodegradation process of phenol by a pure culture of Pseudomonas putida ATCC 17484. The experimental work was carried out in two different systems: a stirred tank where cells grew as a suspended culture and a fluidized bed where cells were immobilized within calcium alginate gel beads. The influence of the hydraulic residence time (HRT) and organic loading rate on the removal efficiency of phenol was determined for both bioreactors. Also, the stability of the fluidized-bed bioreactor (FBB) in terms of its ability to withstand sudden phenol overdoses is also reported. Experimental values indicated that both bioreactors showed high phenol degradation efficiencies, higher than 90%, even for a phenol loading rate in the influent as high as 4 g phenol/l day. The FBB showed better performance than the suspended-culture bioreactor due to its better control and because it could operate with lower HRT.

The inferiorly based rectus abdominis myocutaneous flap for reconstruction of recurrent pressure sores.

Eight neurologically impaired patients underwent reconstruction of chronic perineal and ischial pressure sores utilizing an inferiorly based rectus abdominis myocutaneous flap. Other local and regional flap options had been previously used or were not feasible. In six patients, healing was uncomplicated. One patient required local debridement and flap readvancement. The second involved minor separation of a suture line and healed by secondary intention. All donor sites were closed directly and healed by primary intention. There was no evidence of hernia formation, and no functional deficit was detected from removal of the rectus muscle in any of the patients. In conclusion, it was felt that the inferiorly based rectus abdominis myocutaneous flap should be considered a reconstructive option in dealing with perineal and ischial pressure sores. Furthermore, for reasons discussed, we found distinct advantages to using this flap in spinal cord injury patients.

[Percutaneous treatment of retroperitoneal abscesses]. / Tratamiento percutáneo de los abscesos retroperitoneales.

UNLABELLED: The retroperitoneal abscess is an uncommon disease, that must be treated by drainage. The progressive use of the percutaneous drainage, under ultrasound or computed tomography guidance (CT), has changed the therapeutical management and has demonstrated to be a valid alternative to surgical drainage. From 1986 to 1998, 16 patients with retroperitoneal abscesses were treated by percutaneous drainage (14 with CT and 2 with ultrasound guidance). This method eradicated the abscess in 13 cases, in 2 was necessary a new function to cure the abscess, and 1 patient, with a severe sepsis, died. Percutaneous drainage was the unique treatment used in 12 patients. In the remaining four, the patients' clinical status improved after percutaneous drainage, and they were able to undergo subsequent elective nephrectomy. CONCLUSIONS: Percutaneous drainage of retroperitoneal abscesses has been established as a viable alternative to surgical intervention. This method can resolve the abscess or improve the patient' clinical status to undergo elective surgery.

Deficiency of Kruppel-like factor KLF4 in myeloid-derived suppressor cells inhibits tumor pulmonary metastasis in mice accompanied by decreased fibrocytes.

The importance of immunosuppressive myeloid-derived suppressor cells (MDSCs) bearing monocyte markers in tumor metastasis has been well established. Recently, it was reported that these cells possess phenotypic plasticity and differentiate into fibrocytes, very distinct cells that are precursors of tumorigenic myofibroblasts. However, the importance of this transdifferentiation in tumor metastasis has not been explored. Here, we describe the role of MDSC-derived fibrocytes in tumor metastasis that is regulated by Kruppel-like factor 4 (KLF4), a transcription factor that is critical to monocyte differentiation and to promotion of cancer development. Using mouse metastasis models of melanoma and breast cancer, we found that KLF4 knockout was associated with significantly reduced pulmonary metastasis, which was accompanied by decreased populations of MDSCs, fibrocytes and myofibroblasts in the lung. Cause-effect studies by adoptive transfer revealed that KLF4 deficiency in MDSCs led to significantly reduced lung metastasis that was associated with fewer MDSC-derived fibrocytes and myofibroblasts. Mechanistically, KLF4 deficiency significantly compromised the generation of fibrocytes from MDSCs in vitro. During this process, KLF4 expression levels were tightly linked with those of fibroblast-specific protein-1 (FSP-1), deficiency of which resulted in no metastasis in mice as has been previously reported. In addition, KLF4 bound directly to the FSP-1 promoter as determined by chromatin immunoprecipitation and overexpression of KLF4 increased the FSP-1 promoter activities. Taken together, our results suggest that MDSCs not only execute their immunosuppressive function to promote metastatic seeding as reported before, but also boost metastatic tumor growth after they adopt a fibrocyte fate. Therefore, KLF4-mediated fibrocyte generation from MDSCs may represent a novel mechanism of MDSCs contributing to tumor metastasis and supports the feasibility of inhibiting KLF4 or FSP-1 to prevent tumor metastasis.

Self-perception of health status, mental health and quality of life among adults with diabetes residing in a metropolitan area.

AIM: The study aimed to compare the self-perception of health, physiological distress and health-related quality of life (HRQL) in subjects with and without diabetes residing in a large metropolitan area (the city of Madrid), and to identify the variables associated with the poorest HRQL among diabetes patients. METHODS: In this case-control epidemiological study, we selected 358 patients with diabetes from the Madrid City Health Survey. For every patient, two controls without diabetes were randomly selected from the same database and matched for age, gender and health district. The resultant study population comprised 1074 subjects, who were analyzed according to their self-rated health status, with mental health assessed by the 12-item General Health Questionnaire (GHQ-12) and HRQL by the COOP/WONCA questionnaire. Independent variables included sociodemographic characteristics, lifestyle variables, associated chronic diseases and consumption of medications. Multivariate analyses were conducted using ANCOVA tests. RESULTS: The prevalence of health perceived as fair or poor was 64.12% in those with diabetes vs 38.57% in those without diabetes (P<0.05). The GHQ-12 results showed that mental health was also significantly worse among diabetes sufferers, and the COOP/WONCA questionnaire results indicated significantly poorer HRQL in those with diabetes. The variables that determined a poorer perception of HRQL among diabetes sufferers were female gender, older age, obesity, lack of physical exercise, coexistence of depression, use of sleeping pills, and Alzheimer's and cerebrovascular diseases. CONCLUSION: Self-rated health and psychological well-being, and HRQL, are all considerably poorer among patients with diabetes vs those without diabetes. The poorest quality of life among those with diabetes associated with female gender, depression, lack of exercise and obesity.

The DpsA protein of Synechococcus sp. Strain PCC7942 is a DNA-binding hemoprotein. Linkage of the Dps and bacterioferritin protein families.

The Dps family of proteins are a diverse group of bacterial stress-inducible polypeptides that bind DNA and likely confer resistance to peroxide damage during periods of oxidative stress and long term nutrient limitation. Some members of the Dps protein family have been shown to form large (approximately 150-kDa), hexameric complexes that bind chromosomal DNA with little sequence specificity. In this paper we report the nucleotide sequence of the dpsA gene from Synechococcus sp. PCC7942 encoding a cyanobacterial Dps homolog. The deduced amino acid sequence of the Synechococcus sp. DpsA protein revealed that a carboxyl-terminal domain of the protein was > 60% homologous to the COOH-terminal half of bacterioferritin. Other known Dps family members lack such high similarity to the bacterioferritins. Purification and spectroscopic analysis of the Synechococcus sp. DpsA protein complex revealed that the complex contains heme and has a weak catalase activity in vitro. Activity staining of nondenaturing polyacrylamide gels showed that the protein complex comigrated with both the heme and the catalase activity, and O2 evolution measurements yielded a maximal specific activity of 1.7 mumol of H2O2 consumed/micrograms of protein-1 min-1. We speculate that the protein may have a peroxide-consuming mechanism located on the chromosomal DNA, and we also suggest that this activity may be a necessary feature to handle the endogenous oxidative stresses associated with oxygenic photosynthesis. Last, the evolutionary link between the Dps protein family and the bacterioferritins is discussed.

[Motives for consultation and demographic characteristics of a community of "undocumented" immigrants in the district of Usera-Villaverde (Madrid)]. / Motivos de consulta y características demográficas de una comunidad de inmigrantes "sin papeles" en el distrito de Usera-Villaverde (Madrid).

OBJECTIVE: The objectives of the present study are to describe to the social particularizations, reasons for consultation and diagnoses conducted in the adult immigrants without regularizing that they went to this doctor's office in the district of Villaverde-Usera (Madrid), excluding the data from obstetrics-gynecology and pediatric. DESIGN: One is a descriptive observational study, based on the registry of the daily activity of the consultation from 1996 to 1999. SETTING: Primary level of attention in the area of influence of the municipal districts of Villaverde-Usera. PARTICIPANTS: 1496 consultations to immigrants without regularizing adults, taken care of in our consultation are described in the mentioned period. MEASUREMENTS AND RESULTS: By means of the registry of daily activity social and demographic variables took shelter, of reason and type of consultation, as well as of the main diagnosis that brought to them to the consultation, en 1496 consultations of illegal immigrants (533 people). 31% of the consultations were new and a 14% of absences to the citation were registered. 67% of the consultations made women, the average age was of 34.9 years and in a 76% it referred like South America origin. The diseases that more consultations generated were acute respiratory infection 18%, depressives disorders 11% and the backache with also a 11%. The 48% the reasons for the consultation were acute and 60% took place to free demand. CONCLUSIONS: The group of consultations taken care of responds to the profile of a young South American woman, that fundamentally consults by acute respiratory infections, and very in proximity by depressive-anxious upheavals and lumbar affections, problems very in relation to his situation of immigrant. The irregularity adds to a risk when making difficult the access, most of the consultations are precise, failing in elevated occasions to the programmed consultation.

Purification and characterization of a Synechococcus sp. strain PCC 7942 polypeptide structurally similar to the stress-induced Dps/PexB protein of Escherichia coli.

A stable DNA/protein complex having an apparent molecular mass of approximately 150 kDa was purified from nitrate-limited cultures of the cyanobacterium Synechococcus sp. strain PCC 7942. Amino-terminal peptide sequencing indicated that the polypeptide was structurally similar to the Dps protein of Escherichia coli; Dps is also known as the product of the starvation- and stationary-phase-inducible gene, pexB. The 150-kDa complex dissociated into a 22-kDa protein monomer after boiling in 2% SDS. The 150-kDa complex preparation had approximately a 10% nucleic acid content and upon dissociation released DNA fragments that were sensitive to S1 nuclease digestion. Immunoblot data indicated that the complex accumulates during stationary phase and during nitrogen, sulfur, and phosphorus limitation. DNA-binding assays indicated that the protein nonspecifically binds both linear and supercoiled DNA. Circular dichroism spectroscopy revealed that the Synechococcus sp. Dps-like protein contains extensive regions of alpha-helical secondary structure. We propose that the 150-kDa complex represents a hexameric aggregate of the Dps-like protein complexed with single-stranded DNA and serves to bind a portion of the chromosomal DNA under nutrient-limited conditions.